病毒HIV-1的复制需要病毒组分向免疫细胞的细胞膜开启协调性的运动,而在免疫细胞中病毒粒子就可以装配并且最终释放,近日一篇刊登在国际杂志The Journal of Cell Biology上的研究论文中,来自布宜诺斯艾利斯大学的科学家通过研究揭示了控制胞内运输的蛋白Rab如何通过促进产生高水平的膜脂质来支持HIV-1的装配。在细胞膜特殊位点新装配形成的HIV-1颗粒往往会富含PIP2,PIP2是细胞膜的磷脂组分,其可以补充名为Pr55Gag (Gag)的病毒蛋白,这种病毒蛋白就可以指导HIV-1的装配;目前研究者已经解析了该过程需要的特殊细胞分泌通路,而本文中研究者调查了是否Rab27a蛋白也扮演了类似的功能,该蛋白可以指导内含体向细胞膜的运输。文章中,研究者发现HIV-1的复制在缺失Rab27a蛋白的免疫细胞中处于受损状态,而这些免疫细胞的细胞膜中PIP2的水平也下降了,而且不能够补充Gag蛋白来形成病毒的装配位点;于是研究者就表明,Rab27a蛋白可以通过控制酶类内含体的运输从而促进细胞膜中PIP2的形成,而酶类内含体的运输对于细胞外周脂质的形成非常必要。最后研究者认为,本文的研究成果为后期调查是否控制内含体的运输可以作为一种新型抗HIV-1的疗法提供了新的研究线索,同时也为揭示HIV-1的感染机体提供了一定的帮助
厂商
2015.05.06
表面活性物质关联蛋白D(SPD)检测试剂盒适用生物 Homo sapiens (Human,人) 表面活性物质关联蛋白D(SPD)检测试剂盒 检测范围 6.25-400ng/mL 灵敏度 2.74ng/mL 样本类型 Serum, plasma, tissue homogenates, cell lysates, lung lavage fluid, cell culture supernates and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 表面活性物质关联蛋白D(SPD)检测试剂盒规格 96T 表面活性物质关联蛋白D(SPD)检测试剂盒 ELISA Kit for Surfactant Associated Protein D (SPD)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Homo sapiens (Human) Product No. SEB039Hu Sample type Serum, plasma, tissue homogenates, cell lysates, lung lavage fluid, cell culture supernates and other biological fluids. Format 96-well strip plate Assay length 4.5 hours Detection range 6.25-400ng/mL The standard curve concentrations used for the ELISA’s were 400ng/mL, 200ng/mL, 100ng/mL, 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 2.74ng/mL. 表面活性物质关联蛋白D(SPD)检测试剂盒 SpecificityThis assay has high sensitivity and excellent specificity for detection of Surfactant Associated Protein D (SPD). No significant cross-reactivity or interference between Surfactant Associated Protein D (SPD) and analogues was observed.RecoveryMatrices listed below were spiked with certain level of recombinant Surfactant Associated Protein D (SPD) and the recovery rates were calculated by comparing the measured value to the expected amount of Surfactant Associated Protein D (SPD) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 88-96 92 EDTA plasma(n=5) 79-97 91 heparin plasma(n=5) 78-93 81 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Surfactant Associated Protein D (SPD) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Surfactant Associated Protein D (SPD) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CVLinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Surfactant Associated Protein D (SPD) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 91-99% 97-104% 89-98% 86-103% EDTA plasma(n=5) 85-103% 96-105% 99-105% 92-101% heparin plasma(n=5) 88-95% 92-99% 80-101% 94-102% StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Surfactant Associated Protein D (SPD). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Surfactant Associated Protein D (SPD). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Surfactant Associated Protein D (SPD), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Surfactant Associated Protein D (SPD) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
标准
2015.05.05
谷氨酸半胱氨酸连接酶(GCLM)检测试剂盒适用生物 Homo sapiens (Human,人) 谷氨酸半胱氨酸连接酶(GCLM)检测试剂盒 检测范围 0.156-10ng/mL 灵敏度 0.061ng/mL 样本类型 Tissue homogenates and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 谷氨酸半胱氨酸连接酶(GCLM)检测试剂盒规格 96T ELISA Kit for Glutamate Cysteine Ligase, Modifier Subunit (GCLM)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!谷氨酸半胱氨酸连接酶(GCLM)检测试剂盒 Organism species Homo sapiens (Human) Product No. SEB038Hu Sample type Tissue homogenates and other biological fluids. Format 96-well strip plate Assay length 4.5 hours Detection range 0.156-10ng/mL The standard curve concentrations used for the ELISA’s were 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.313ng/mL, 0.156ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 0.061ng/mL. 谷氨酸半胱氨酸连接酶(GCLM)检测试剂盒 SpecificityThis assay has high sensitivity and excellent specificity for detection of Glutamate Cysteine Ligase, Modifier Subunit (GCLM). No significant cross-reactivity or interference between Glutamate Cysteine Ligase, Modifier Subunit (GCLM) and analogues was observed.PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glutamate Cysteine Ligase, Modifier Subunit (GCLM) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glutamate Cysteine Ligase, Modifier Subunit (GCLM) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CVStabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Glutamate Cysteine Ligase, Modifier Subunit (GCLM). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Glutamate Cysteine Ligase, Modifier Subunit (GCLM). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Glutamate Cysteine Ligase, Modifier Subunit (GCLM), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Glutamate Cysteine Ligase, Modifier Subunit (GCLM) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
标准
2015.05.05
补体成分4(C4)检测试剂盒适用生物 Homo sapiens (Human,人) 检测范围 9.38-600ng/mL 灵敏度 4.3ng/mL 样本类型 Serum, plasma and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 规格 96T 补体成分4(C4)检测试剂盒ELISA Kit for Complement Component 4 (C4)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Homo sapiens (Human) Product No. SEA888Hu Sample type Serum, plasma and other biological fluids. Format 96-well strip plate Assay length 4.5 hours Detection range 9.38-600ng/mL The standard curve concentrations used for the ELISA’s were 600ng/mL, 300ng/mL, 150ng/mL, 75ng/mL, 37.5ng/mL, 18.75ng/mL, 9.38ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 4.3ng/mL. SpecificityThis assay has high sensitivity and excellent specificity for detection of Complement Component 4 (C4). No significant cross-reactivity or interference between Complement Component 4 (C4) and analogues was observed.补体成分4(C4)检测试剂盒RecoveryMatrices listed below were spiked with certain level of recombinant Complement Component 4 (C4) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 4 (C4) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 97-105 101 EDTA plasma(n=5) 95-102 98 heparin plasma(n=5) 87-101 91 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 4 (C4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Complement Component 4 (C4) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CVLinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Complement Component 4 (C4) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 96-103% 87-103% 78-92% 89-97% EDTA plasma(n=5) 81-99% 86-98% 87-96% 87-94% heparin plasma(n=5) 89-102% 78-89% 81-104% 85-99% 补体成分4(C4)检测试剂盒StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. 补体成分4(C4)检测试剂盒Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Complement Component 4 (C4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Complement Component 4 (C4). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Complement Component 4 (C4), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Complement Component 4 (C4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
标准
2015.05.04
肾素(REN)检测试剂盒适用生物 Homo sapiens (Human,人) 检测范围 31.25-2000pg/mL 灵敏度 13.3pg/mL 样本类型 Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 规格 96T 肾素(REN)检测试剂盒ELISA Kit for Renin (REN)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Homo sapiens (Human) Product No. SEA889Hu Sample type Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Format 96-well strip plate Assay length 4.5 hours Detection range 31.25-2000pg/mL The standard curve concentrations used for the ELISA’s were 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL Sensitivity The minimum detectable dose of this kit is typically less than 13.3pg/mL. 肾素(REN)检测试剂盒SpecificityThis assay has high sensitivity and excellent specificity for detection of Renin (REN). No significant cross-reactivity or interference between Renin (REN) and analogues was observed.RecoveryMatrices listed below were spiked with certain level of recombinant Renin (REN) and the recovery rates were calculated by comparing the measured value to the expected amount of Renin (REN) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 80-88 84 EDTA plasma(n=5) 81-101 97 heparin plasma(n=5) 95-103 101 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Renin (REN) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Renin (REN) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CV肾素(REN)检测试剂盒LinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Renin (REN) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 79-90% 80-104% 86-102% 85-98% EDTA plasma(n=5) 93-105% 85-101% 90-98% 87-94% heparin plasma(n=5) 90-103% 90-97% 99-105% 98-105% StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. 肾素(REN)检测试剂盒Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Renin (REN). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Renin (REN). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Renin (REN), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Renin (REN) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
标准
2015.05.04
美国芝加哥大学科学家在《免疫》期刊上撰文指出,身体内的免疫系统可能是健康肠道菌群“卫士”。他们发现,白血球中的一种单一结合蛋白质可能影响小鼠的肠道菌群是否平衡。如果没有该蛋白质,小鼠更容易感染有害细菌。但其背后的机制尚不清楚,科学家表示,可能是免疫系统能以某种方式感知到入侵肠道细菌的存在。“我们的研究揭示了人体免疫系统赋予肠道细菌天然的限制感染能力。”芝加哥大学病理学系yang-xinfu说,“由于耐抗生素的有害细菌急剧增长,科学家亟须找到方法,在不使用抗生素的前提下,控制有害细菌感染。未来,这种方法或能让有益细菌间接杀死有害细菌。”fu及其合作者发现,当缺少一种名为id2的蛋白质时,一种名为固有淋巴细胞3型的肠道免疫细胞(ilc3)对有害细菌感染的响应能力就会减弱。缺乏id2的ilc3无法产生一种名为il-22的分子,该分子能刺激其他肠道细胞产生抗菌的缩氨酸(amp),从而帮助身体抵抗病原菌感染。但正常细菌似乎对amp具有更强的抵抗力
百态
2015.05.04
在某些种类的癌症中,神经和癌症细胞会跳一种常常致命且复杂的华尔兹,癌症细胞和神经相互靠近,最终,癌症细胞进入到了神经中。这项发表在Nature Communications杂志上的研究,对传统的关于周围神经浸润的观点发起了挑战。传统的观点认为癌症细胞是侵略者,它们通过阻力最小的路径入侵神经,密歇根大学牙科院教授和首席研究员Nisha D'Silva说。D'Silva的实验室发现,外周神经浸润事实上是,在神经和癌症细胞之间的,更为复杂的,经过编排的生物化学的取与舍。"一旦头颈部肿瘤侵入到了神经,这是可能发生的最糟糕的事情之一," D'Silva说。她也在U-M医学院病理系任职,是U-M癌症中心头颈肿瘤项目的成员之一。"癌症细胞侵入神经,与患者的低存活率高度相关,目前仍没有靶向治疗药物,因为我们并不清楚,为什么有些肿瘤细胞这样做,有些不这样做。"外周神经浸润常见于,头颈,胰腺,胃部和肠道肿瘤,引起严重的疼痛和麻木,肿瘤扩散和复发,功能丧失,和其它的一些并发症。D'Silva的实验室发现,外周神经浸润起始于神经细胞释放一种刺激,从而在肿瘤细胞上激发了一种特殊的蛋白受体。这种受体在肿瘤细胞激活了指令,反向给神经细胞释放同样的刺激。神经辨认出了这种刺激,从而向肿瘤细胞前进。想象一下,两个舞者在一个房间认出了对方,并缓慢地向对方靠近,直到他们成为了永久的伴侣。在起始的配对后,这种信号环路一直持续着。"基本上,它们是在跳华尔兹," D'Silva说,"这是一支十分优雅的舞蹈,如果你愿意想象的话。"在头颈肿瘤研究外周神经浸润是非常困难的,所以,D'Silva的实验室不得不开发出一种方法在活样本中观察这种相互作用。首先,研究人员将神经移植到鸡蛋膜上,等神经融合后,他们再研究神经和头颈肿瘤之间的相互作用。D'Silva说下一步的研究是, "我们什么时候和如何来解释这种舞蹈。"这项研究题目是"甘丙肽调节神经环境以有益于头颈肿瘤的外周神经浸润
厂商
2015.05.04
血管紧张素原(AGT)检测试剂盒适用生物 Homo sapiens (Human,人) 检测范围 1.56-100ng/mL 灵敏度 0.72ng/mL 样本类型 Serum, plasma, urine, tissue homogenates, cell lysates, cell culture supernates and other biological fluid. 实验时长 4.5h 实验方法 双抗夹心法 规格 96T血管紧张素原(AGT)检测试剂盒 ELISA Kit for Angiotensinogen (AGT)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Homo sapiens (Human) Product No. SEA797Hu Sample type Serum, plasma, urine, tissue homogenates, cell lysates, cell culture supernates and other biological fluid. Format 96-well strip plate Assay length 4.5 hours Detection range 1.56-100ng/mL The standard curve concentrations used for the ELISA’s were 100ng/mL, 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL, 3.13ng/mL, 1.56ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 0.72ng/mL. SpecificityThis assay has high sensitivity and excellent specificity for detection of Angiotensinogen (AGT). No significant cross-reactivity or interference between Angiotensinogen (AGT) and analogues was observed.血管紧张素原(AGT)检测试剂盒 RecoveryMatrices listed below were spiked with certain level of recombinant Angiotensinogen (AGT) and the recovery rates were calculated by comparing the measured value to the expected amount of Angiotensinogen (AGT) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 88-99 94 EDTA plasma(n=5) 88-95 92 heparin plasma(n=5) 87-103 98 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Angiotensinogen (AGT) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Angiotensinogen (AGT) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CVLinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Angiotensinogen (AGT) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 93-102% 93-101% 98-105% 82-104% EDTA plasma(n=5) 95-103% 82-93% 92-101% 97-105% heparin plasma(n=5) 84-98% 88-96% 79-102% 98-105% 血管紧张素原(AGT)检测试剂盒 StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Angiotensinogen (AGT). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Angiotensinogen (AGT). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Angiotensinogen (AGT), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Angiotensinogen (AGT) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
标准
2015.04.30
尊敬的客户,公司各职员: 您们好!感谢大家长期关注上海钰博生物动态,根据国家法定假期的规定,并结合公司实际情况,现对五一节放假做如下安排:一、放假时间5月1日至3日(星期五至星期天)放假,共3天。其中,5月1日(星期五)为五一假期,5月2日(星期六),5月3日(星期天)为法定节假日照常公休。二、请公司各职员做好自己的节前工作安排。三、放假期间本公司不安排人员值班,如有订购产品请发送留言至我司网站平台,我们会在节后上班第一时间及时为您办理订货发货。各网站均有在线留言咨询栏。对此给您带来的不便我们深感抱歉。感谢你的支持与理解。 上海钰博 特此通知!
标准
2015.04.30
半胱氨酸蛋白酶抑制剂3(CST3)检测试剂盒适用生物 Homo sapiens (Human,人) 检测范围 1.56-100ng/mL 灵敏度 0.67ng/mL 样本类型 Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 规格 96T 半胱氨酸蛋白酶抑制剂3(CST3)检测试剂盒 ELISA Kit for Cystatin 3 (CST3)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Homo sapiens (Human) Product No. SEA896Hu Sample type Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Format 96-well strip plate Assay length 4.5 hours Detection range 1.56-100ng/mL The standard curve concentrations used for the ELISA’s were 100ng/mL, 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL, 3.13ng/mL, 1.56ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 0.67ng/mL.半胱氨酸蛋白酶抑制剂3(CST3)检测试剂盒 SpecificityThis assay has high sensitivity and excellent specificity for detection of Cystatin 3 (CST3). No significant cross-reactivity or interference between Cystatin 3 (CST3) and analogues was observed. RecoveryMatrices listed below were spiked with certain level of recombinant Cystatin 3 (CST3) and the recovery rates were calculated by comparing the measured value to the expected amount of Cystatin 3 (CST3) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 98-105 102 EDTA plasma(n=5) 92-101 97 heparin plasma(n=5) 79-101 94 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cystatin 3 (CST3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cystatin 3 (CST3) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CV半胱氨酸蛋白酶抑制剂3(CST3)检测试剂盒 LinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cystatin 3 (CST3) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 81-93% 89-101% 93-101% 84-98% EDTA plasma(n=5) 84-98% 95-105% 93-104% 84-99% heparin plasma(n=5) 93-101% 98-105% 79-99% 81-99% StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end. Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Cystatin 3 (CST3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Cystatin 3 (CST3). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Cystatin 3 (CST3), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Cystatin 3 (CST3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
厂商
2015.04.29
铜蓝蛋白(CP)检测试剂盒适用生物 Homo sapiens (Human,人) 检测范围 6.25-400ng/mL 灵敏度 2.54ng/mL 样本类型 Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 规格 96T 铜蓝蛋白(CP)检测试剂盒 ELISA Kit for Ceruloplasmin (CP)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Homo sapiens (Human) Product No. SEA909Hu Sample type Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Format 96-well strip plate Assay length 4.5 hours Detection range 6.25-400ng/mL The standard curve concentrations used for the ELISA’s were 400ng/mL, 200ng/mL, 100ng/mL, 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 2.54ng/mL.铜蓝蛋白(CP)检测试剂盒 SpecificityThis assay has high sensitivity and excellent specificity for detection of Ceruloplasmin (CP). No significant cross-reactivity or interference between Ceruloplasmin (CP) and analogues was observed. RecoveryMatrices listed below were spiked with certain level of recombinant Ceruloplasmin (CP) and the recovery rates were calculated by comparing the measured value to the expected amount of Ceruloplasmin (CP) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 78-97 87 EDTA plasma(n=5) 99-105 102 heparin plasma(n=5) 94-102 98 铜蓝蛋白(CP)检测试剂盒 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ceruloplasmin (CP) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Ceruloplasmin (CP) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CVLinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Ceruloplasmin (CP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 90-98% 94-101% 93-101% 99-105% EDTA plasma(n=5) 88-101% 80-95% 79-102% 91-98% heparin plasma(n=5) 80-98% 92-99% 88-101% 86-98% StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end. Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Ceruloplasmin (CP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Ceruloplasmin (CP). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Ceruloplasmin (CP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Ceruloplasmin (CP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
厂商
2015.04.29
尽管单一的免疫检验点抑制剂作为肿瘤免疫治疗,显示出了的好的前景和希望。然而,最近有数据显示,将这些抑制剂联合使用,可能再加上其他的治疗方法,例如,放疗,将可以取得更大的成功。Nature杂志上报道,Minn和其同事们发现,联合抗体(分别靶向细胞程序性死亡蛋白1(PD1)配体1(PDL1)介导的和细胞毒性T淋巴细胞抗原4(CTLA4)介导的免疫检验点)和放疗,可以通过不同的,非重复性的机制,有效地促进抗肿瘤免疫。在22个转移性黑色素瘤病人的一期临床试验中,一个单一的病变在照射后,采用了4轮CTLA4特异性单克隆抗体ipilimumab的治疗。通过对非照射的黑色素瘤病变的分析,研究人员们发现,18%的病人有部分反应,另有18%的病人在治疗后病情稳定,这些结果提示,在一部分病人中,放疗和ipilimumab的联合疗法有有益的影响。然而,64%的病人是对治疗抵抗的,病情仍在发展中。为了研究清楚肿瘤的耐药机制,研究人员们利用B6-F10黑色素瘤老鼠模型,在使用放疗和CTLA抑制剂联合治疗后,他们观察到了类似的反应比例。完全缓解疗法是依赖于CD8+ T细胞的,且最好的肿瘤耐药的预报是,在肿瘤浸润淋巴细胞群中(TIL),CD8+CD44+ T细胞和调节性T细胞(TReg)的比例(CD8/TReg ratio)升高失败。尽管TReg 细胞的数量有所降低(在敏感性细胞中也有类似的发现),但CD8/TReg的比例并没有升高,这是因为CD8+ T 细胞没有累积。在耐药肿瘤中,CD8+ T 细胞累积的缺陷是由于在黑色素瘤上PDL1的高表达引起的。在耐药的乳腺癌细胞上和临床试验中,来源于转移性多发黑色素瘤病人的耐药细胞上,均可以发现一个相似的PDL1的高表达。增高的PDL1可以促进T细胞的消耗(通过检测PD1+EOMES+CD8+ T 细胞中的Ki67和颗粒酶B的表达可知),其逆转可称为复兴。在敏感性细胞,放疗和CTLA抑制剂联合治疗,可以升高复兴细胞与消耗掉的CD8+ T细胞在TIL群中的比例。然而,在耐药细胞,复兴的CD8+ T细胞数目并没有增高。但是,额外再添加PDL1特异性阻断抗体后,复兴的CD8+ TILs和CD8/TReg均有所增加。重要的是,黑色素瘤老鼠的完全缓解率从双联合治疗的17%提高到了三项联合治疗的80%,即CTLA和PDL1(PD1)阻断加放疗。进一步的分析表明,这三项的联合治疗是一种非重复的,互补的方式:PDL1阻断疗法复兴了耗尽的CD8+ T细胞,CTLA4阻断疗法主要减少了TReg 的细胞数量,两者加在一起,这些免疫检验点抑制剂增加了CD8/TReg 的比例,促进了TILs细胞的四周克隆扩增。放疗的主要作用是多元化TILs的T细胞受体,塑造出扩增的外周细胞克隆。值得注意的是,外周T细胞消耗和复兴的标志,血液中CD8/TReg 的比例,可用来预测老鼠对治疗的反应性。而且,在临床试验的病人中,预处理肿瘤活检黑色瘤细胞中PDL1的高表达,与放疗和ipilimumab 联合治疗后,持续的T细胞消耗,快速的疾病发展相关。这些数据表明,肿瘤细胞上PDL1的高表达,是肿瘤细胞对放疗加CTLA4阻断剂治疗的主导抵抗机制。这些数据支持探索一种治疗方法,即将阻断PDL1(或PD1)和CTLA及放疗联合起来,治疗黑色素瘤和其他可能的实体瘤
百态
2015.04.29
你也许遗传了你妈妈美丽的眼神,但同时她也给了你线粒体的DNA突变,所谓母系遗传疾病的根源。一项基于小鼠的实验表示可以通过两种技术大幅降低卵子中有害DNA的风险,从而使子女能够逃避遗传类的疾病。此种方法也规避了存在伦理问题的"线粒体置换技术"-该技术会导致"三亲"型的胚胎。尽管研究人员没有在人类胚胎中对此项技术进行试验,然而此项技术确实是"前所未有的"。来自美国南阿拉巴马大学的分子生物学家Mikhail Alexeyev认为此项试验是第一次在受精卵中进行的。线粒体是细胞内产生能量的场所,这种细胞器含有自身的DNA,与细胞核内的大部分遗传物质相互隔离,但这并不是它们唯一的奇特之处。由于受精卵中的线粒体只能来自于卵子而非精子,所以线粒体是从母亲遗传下来的细胞器。然而,不幸的是没200个女性中就有一个携带缺陷型的线粒体DNA,她们的子女从而会得一系列致死性的疾病,比如周期性呕吐综合征、Leigh综合征等。线粒体疾病是十分罕见,同时也是难以治愈的疾病。研究者们试图开发不同的方法去除线粒体中受损的DNA,或者阻止携带此类突变DNA的母亲将致病性线粒体传递给下一代的子女。其中一类叫做线粒体置换技术的手段,通过将代孕母亲卵子中细胞核转移至正常女子的去核卵细胞中,重组后的卵子再经过体外受精之后转移至代孕母亲的体内。然而,这一方法受到了多方面的质疑。科学家们怀疑该方法的安全性,而伦理学家认为该方法导致出生后的婴儿携带三个亲本的遗传物质。今年年初,英国批准了这一治疗方法,而美国FDA仍在考虑之中。除了将坏掉的线粒体用好的进行置换之外,美国沙克研究所的发育学家Juan Carlos Izpisua Belmonte带领其团队研究如何直接改造线粒体中的DNA。研究者们首先测试了将一条带有DNA剪切酶翻译序列以及线粒体定位序列的RNA链导入受体细胞中,该RNA链进入线粒体之后能够产生DNA剪切酶,从而将损坏的线粒体DNA进行剪切,Izpisua Belmonte与同事们将RNA导入线粒体DNA具有特定突变的小鼠卵子或早期的胚胎中,之后检测该方法在突变DNA清除方面的效果。相关结果发表在今天的《cell》期刊上。为了研究该方法是否会对胚胎造成损伤,研究者们将一些处理后的胚胎导入雌性小鼠中,这些小鼠均生产出了健康的后代,而且后代的小鼠也可以正常发育。在这一研究中,针对的DNA靶向序列其实是不致病的,但是这项研究的研究依然说明这一技术可以用于以上的疾病治疗。限制性内切酶技术存在一定的局限性。研究者们发现该技术只对两类线粒体疾突变中的一种有效果。因而他们又研究了另外一类基因编辑技术-TALEN,从而更加自由的对线粒体DNA进行编辑。当研究者们改用TALEN技术进行线粒体DNA编辑之后,发现仍然只能降低其中一种类型的突变量。然而当改用在人类或小鼠疾病相关线粒体DNA突变的治疗试验中时,作者们发现可以降低两类疾病相关的线粒体DNA突变。尽管在生殖科的临床治疗方面已经有运用RNA注射方法的历史,然而没有针对消除卵子或受精卵中靶向DNA的前例。大多数人细胞中都有一定程度的线粒体DNA突变,当这一突变比例上升到60%到95%时就会产生相关的症状。我们要做的是将这一比例降低到症状显现的阈值以下
新品
2015.04.29
甲胎蛋白(αFP)检测试剂盒适用生物 Rattus norvegicus (Rat,大鼠) 检测范围 0.313-20ng/mL 灵敏度 0.119ng/mL 样本类型 Serum, plasma, tissue homogenates and other biological fluids 实验时长 4.5h 实验方法 双抗夹心法 规格 96T ELISA Kit for Alpha-Fetoprotein (aFP)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Rattus norvegicus (Rat) Product No. SEA153Ra Sample type Serum, plasma, tissue homogenates and other biological fluids Format 96-well strip plate Assay length 4.5 hours Detection range 0.313-20ng/mL The standard curve concentrations used for the ELISA’s were 20ng/mL, 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.313ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 0.119ng/mL. SpecificityThis assay has high sensitivity and excellent specificity for detection of Alpha-Fetoprotein (aFP). No significant cross-reactivity or interference between Alpha-Fetoprotein (aFP) and analogues was observed.RecoveryMatrices listed below were spiked with certain level of recombinant Alpha-Fetoprotein (aFP) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-Fetoprotein (aFP) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 85-92 89 EDTA plasma(n=5) 80-101 81 heparin plasma(n=5) 91-104 98 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Alpha-Fetoprotein (aFP) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-Fetoprotein (aFP) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CVLinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Alpha-Fetoprotein (aFP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 90-104% 78-101% 89-103% 78-98% EDTA plasma(n=5) 81-99% 82-99% 96-104% 81-90% heparin plasma(n=5) 88-105% 80-90% 95-102% 98-105% StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Alpha-Fetoprotein (aFP). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Alpha-Fetoprotein (aFP). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Alpha-Fetoprotein (aFP), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Alpha-Fetoprotein (aFP) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
厂商
2015.04.28
激肽释放酶3(KLK3)检测试剂盒适用生物 Rattus norvegicus (Rat,大鼠) 检测范围 0.156-10ng/mL 灵敏度 0.063ng/mL 样本类型 Serum, plasma, tissue homogenates and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 规格 96TELISA Kit for Kallikrein 3 (KLK3)FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!Organism species Rattus norvegicus (Rat) Product No. SEA151Ra Sample type Serum, plasma, tissue homogenates and other biological fluids. Format 96-well strip plate Assay length 4.5 hours Detection range 0.156-10ng/mL The standard curve concentrations used for the ELISA’s were 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.313ng/mL, 0.156ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 0.063ng/mL. SpecificityThis assay has high sensitivity and excellent specificity for detection of Kallikrein 3 (KLK3). No significant cross-reactivity or interference between Kallikrein 3 (KLK3) and analogues was observed.RecoveryMatrices listed below were spiked with certain level of recombinant Kallikrein 3 (KLK3) and the recovery rates were calculated by comparing the measured value to the expected amount of Kallikrein 3 (KLK3) in samples. Matrix Recovery range (%) Average(%) serum(n=5) 99-105 102 EDTA plasma(n=5) 89-101 92 heparin plasma(n=5) 82-91 86 PrecisionIntra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Kallikrein 3 (KLK3) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Kallikrein 3 (KLK3) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVInter-Assay: CVLinearityThe linearity of the kit was assayed by testing samples spiked with appropriate concentration of Kallikrein 3 (KLK3) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Sample 1:2 1:4 1:8 1:16 serum(n=5) 80-99% 84-101% 84-98% 93-105% EDTA plasma(n=5) 88-98% 87-101% 79-104% 90-98% heparin plasma(n=5) 80-90% 83-97% 87-97% 95-102% StabilityThe stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.Reagents and materials providedReagents Quantity Reagents Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard 2 Standard Diluent 1×20mL Detection Reagent A 1×120μL Assay Diluent A 1×12mL Detection Reagent B 1×120μL Assay Diluent B 1×12mL TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1 Assay procedure summary1. Prepare all reagents, samples and standards;2. Add 100μL standard or sample to each well. Incubate 2 hours at 37oC;3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;4. Aspirate and wash 3 times;5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;6. Aspirate and wash 5 times;7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;8. Add 50μL Stop Solution. Read at 450nm immediately. Test principleThe test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Kallikrein 3 (KLK3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Kallikrein 3 (KLK3). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Kallikrein 3 (KLK3), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Kallikrein 3 (KLK3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
厂商
2015.04.28
粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒适用生物 Homo sapiens (Human,人) 检测范围 31.25-2000pg/mL 灵敏度 12.6pg/mL 样本类型 Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. 实验时长 4.5h 实验方法 双抗夹心法 规格 96T 粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒(酶联免疫吸附试验法)适用生物:人使用说明书仅供体外研究使用,不用于临床诊断!粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒[预期应用]本试剂盒运用双抗体夹心ELISA 法定量测定人血清、血浆、组织匀浆、细胞裂解液、细胞培养上清或其它相关生物液体中GMCSF 含量。[试剂盒内容]试剂名称数量试剂名称数量96孔板(预包被) 1 96孔板覆膜4标准品2 标准品稀释液1×20mL检测溶液A 1×120μL 检测稀释液A 1×12mL检测溶液B 1×120μL 检测稀释液B 1×12mLTMB底物1×9mL 终止液1×6mL浓洗涤液(30×) 1×20mL 使用说明书1[需自备的设备及试剂]1、450±10nm 滤光片的酶标仪(建议仪器使用前提前预热)2、单道或多道微量加液器及吸头3、稀释样品的EP 管4、蒸馏水或去离子水5、吸水纸6、盛放洗液的容器[试剂盒的储存及有效期]1、未开封的试剂盒:所有试剂均按试剂瓶标签上所示保存。请注意,收到试剂盒后请尽快将标准品、检测溶液A、检测溶液B 以及96 孔板保存于-20oC。2、使用后的试剂盒:剩余试剂仍需按照试剂瓶标签所示的温度保存,开封后的酶标板要加干燥剂后密封保存于-20oC,避免潮湿。注意:试剂盒内酶标条可拆卸,按实验需求可分多次使用;使用后的剩余试剂盒建议在首次实验后1 个月内使用完毕。产品过期时间以盒子上的标签为准,保质期内所有组分都确保是稳定的。[标本的采集与保存]1、血清:将收集于血清分离管的全血标本在室温放置2 小时或4oC 过夜,然后1000×g 离心20 分钟,取上清即可,或将上清置于-20oC 或-80oC 保存,但应避免反复冻融。2、血浆:用EDTA 或肝素作为抗凝剂采集标本,并将标本在采集后的30 分钟内于2-8oC 1000×g 离心15 分钟,取上清即可检测,或将上清置于-20oC 或-80oC 保存,但应避免反复冻融。3、组织匀浆:1) 取适量组织块,于预冷PBS(0.01mol/L, pH 7.0-7.2)中清洗去除血液,称重后备用(组织块较大需先剪碎后再匀浆);2) 可同时选用多种匀浆方法达到较好的破碎效果:首先将组织块移入玻璃匀浆器,加入5-10mL 预冷PBS 进行充分研磨,该过程需在冰上进行(有条件实验室可选用机器匀浆);得到的匀浆液可再利用超声破碎或反复冻融进一步处理(超声破碎过程中注意冰浴降温;反复冻融法可重复2 次)。3) 将制备好的匀浆液于5000×g 离心5 分钟,留取上清即可检测。4、细胞裂解液:1)贴壁细胞需要先用胰酶消化,离心收集细胞(悬浮细胞可直接离心收集);2)将收集到的细胞用冷PBS 洗3 次;3)物理方法裂解细胞(可先超声破碎细胞,再反复冻融):i 超声破碎:取适量PBS 重悬细胞,用一定功率的超声波处理细胞悬液,使细胞急剧震荡破裂。ii 反复冻融:将待破碎的细胞在-20oC 以下冰冻,室温融解,反复3 次,使细胞溶胀破碎。4)将标本于2-8oC 1500×g 离心10 分钟,收集上清备用。5、细胞培养上清或其它生物标本:请1000×g 离心20 分钟,取上清即可检测,或将上清置于-20oC 或-80oC 保存,但应避免反复冻融。注意:1、以上标本均需密封保存,4oC 保存应小于1 周,-20oC 不应超过1 个月,-80oC 不应超过2 个月。2、标本溶血会影响最后检测结果,因此溶血标本不宜进行此项检测。3、标本使用前应缓慢均衡至室温,不应加热使之融解。[试剂准备]1、使用前将所有的试剂和标本缓慢均衡至室温(18-25oC),试剂不能直接在37oC 溶解。2、标准品(冻干品):每瓶标准品加入标准品稀释液1mL,盖好后室温静置大约10 分钟,同时反复颠倒/搓动以助溶解,其浓度为4,000pg/mL(贮液)。先将其稀释为2,000pg/mL(标准曲线最高浓度)后,再准备7 个稀释标准品的EP 管,每个EP 管中加入500μL 的标准品稀释液,如图所示依次倍比稀释成2,000pg/mL,1,000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.2pg/mL,标准品稀释液(0pg/mL)直接作为空白孔。为保证实验结果有效性,每次实验请使用新的标准品溶液。Tube 1 2 3 4 5 6 7 8 9pg/mL 4,000 2,000 1,000 500 250 125 62.5 31.2 03、检测溶液A 及检测溶液B:Detection A 及Detection B 在使用前请手甩几下或少时离心处理,以使管壁或瓶盖的液体沉积到管底。临用前分别以检测稀释液A 或B1:100 稀释(如:10μL 检测溶液A/990μL 检测稀释液A),充分混匀,稀释前根据预先计算好的每次实验所需的总量配制(100μL/孔),实际配制时应多配制0.1-0.2mL。4、浓洗涤液:用580mL 蒸馏水或去离子水将20mL 浓洗涤液稀释至600mL,进行30 倍稀释。5、底物溶液:请用灭菌的移液器吸头吸取所需体积的TMB 至另一干净容器中使用,容器中剩余的底物应予丢弃,不要倒回TMB 瓶中。注意:1、标准品的稀释不能直接在板中进行。2、标准品请于临用前15 分钟内配制。该标准品只能使用一次。3、标准品、检测溶液A 工作液、检测溶液B 工作液请使用相应的稀释液配制,不能混淆。混匀时要轻轻充分混匀,避免起泡。为保证实验结果的准确请使用微量吸管,并校准微量加液器。请依据所需的量精确配制,尽量不要微量配制(如吸取检测溶液A 时,一次不要小于10μL),以避免造成浓度误差。4、请勿重复使用已经稀释过的标准品、检测溶液A 工作液和检测溶液B 工作液。5、浓洗涤液中如有结晶析出,请先温育至室温,轻轻混匀,至到结晶完全溶解再进行配制。6、试剂盒中部分试剂为储存液,客户需配置成工作液后使用,在配置过程中如因纯净水质量差或水质污染,以及实验中所用耗材洁净度差,可能造成实验结果不准确,甚至完全错误,请使用双蒸水。[标本处理]1、本公司只对试剂盒本身负责,不对因使用该试剂盒所造成的样本消耗负责,请使用者使用前充分考虑到样本的可能使用量,预留充足的样本。2、实验前应预测标本含量,如果标本浓度过高,应对标本进行稀释,使稀释后的标本符合试剂盒的检测范围,计算时再乘以相应的稀释倍数。标本使用0.01mol/L 的PBS 稀释(PH=7.0-7.2)。3、若所检样本不包含在说明书所列样本之中,建议进行预实验验证其有效性,并注意留存样本。4、使用化学裂解液制备的组织匀浆或细胞提取液可能会由于某些化学物质的引入导致ELISA 实验结果偏差。5、若样本为细胞培养上清,因该类样本干扰因素较多,如:细胞状态、细胞数量、采样时间等,所以可能存在检测不出的情况。6、某些天然蛋白或重组蛋白,包括原核及真核重组蛋白,可能因为与本产品所使用的检测抗体及捕获抗体不匹配,而不被检测出。7、建议使用新鲜样本,保存时间过长可能会存在蛋白降解或变性导致实验结果偏差。[操作步骤]1、加样:分别设标准孔、待测样品孔、空白孔。设标准孔7 孔,依次加入100μL 不同浓度的标准品(见试剂准备2)。空白孔加100μL(见试剂准备第二步最后一管),余孔加待测样品100μL,酶标板加上覆膜,37oC 温育2 小时。2、弃去液体,甩干,不用洗涤。3、每孔加检测溶液A 工作液100μL(临用前配制),酶标板加上覆膜,37oC 温育1 小时。4、弃去孔内液体,每孔用350μL 的洗涤液洗涤,浸泡1-2 分钟,吸去(不可触及板壁)或甩掉酶标板内的液体,在实验台上铺垫几层吸水纸,酶标板朝下用力拍几次(也可轻拍将孔内液体拍干),重复洗板3 次。最后一次洗涤后,要把孔内的洗涤液完全甩干。自动洗板机亦可。5、每孔加检测溶液B 工作液(临用前配制)100μL,加上覆膜,37oC 温育30 分钟。6、弃去孔内液体,甩干,洗板5 次,方法同步骤4。7、每孔加底物溶液90μL,酶标板加上覆膜,37oC 避光显色(反应时间控制在15-25 分钟,不要超过30 分钟。当标准孔的前3-4 孔有明显的梯度蓝色,后3-4 孔梯度不明显时,即可终止)。8、每孔加终止溶液50μL,终止反应,此时蓝色立转黄色。终止液的加入顺序应尽量与底物溶液的加入顺序相同。如出现颜色不匀一,请轻轻晃动酶标板以使溶液混合均匀。9、在确保酶标板底无水滴及孔内无气泡后,立即用酶标仪在450nm 波长测量各孔的光密度(O.D.值)。注意:1、试剂准备:准备一次实验所需要的酶标条,其它的可从微孔板上拆下,密封,按照说明书要求保存,以备下次使用。2、加样:实验操作中请使用一次性的吸头,避免交叉污染。加样时注意不要有气泡,将样品加于酶标板底部,尽量不触及孔壁,轻轻晃动混匀。加样或加试剂时,第一个孔与最后一个孔加样之间的时间间隔如果太大,将会导致不同的“预温育”时间,从而明显地影响到测量值的准确性及重复性。因此,一次加样时间(包括标准品及所有样品)最好控制在10 分钟内。推荐设置复孔进行实验。3、温育:为防止样品蒸发,实验时请将加上盖或覆膜的酶标板置于湿盒内,以避免液体蒸发,洗板后应尽快进行下步操作,任何时侯都应避免酶标板处于干燥状态,同时应严格遵守给定的温育时间和温度。4、洗涤:充分的洗涤非常重要,在每次洗涤过程中,都要将洗涤液完全甩干。洗涤过程中反应孔中残留的洗涤液应在滤纸上拍干,勿将滤纸直接放入反应孔中吸水,同时要消除板底残留的液体和手指印,避免影响最后的酶标仪读数。如果有自动洗板机,应在熟练使用后再用到正式实验过程中。5、反应时间的控制:加入底物后请定时观察反应孔的颜色变化(比如,每隔10 分钟观察一次),如颜色较深,请提前加入终止液终止反应,避免反应过强从而影响酶标仪光密度读数。6、底物:底物请避光保存,在储存和温育时避免强光直接照射。7、如果实验室内湿度低于60%,推荐使用加湿器提高湿度水平。[实验原理]将GMCSF 抗体包被于96 孔微孔板中,制成固相载体,向微孔中分别加入标准品或标本,其中的GMCSF 与连接于固相载体上的抗体结合,然后加入生物素化的GMCSF 抗体,将未结合的生物素化抗体洗净后,加入HRP 标记的亲和素,再次彻底洗涤后加入TMB 底物显色。TMB 在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的GMCSF 呈正相关。用酶标仪在450nm 波长下测定吸光度(O.D.值),计算样品浓度。[计算]各标准品及样本O.D.值扣除空白孔O.D.值后作图(七点图),如设置复孔,则应取其平均值计算。以标准品的浓度为纵坐标(或对数坐标),O.D.值为横坐标(或对数坐标),绘出标准曲线(最佳方程式应依回归方程计算的R2值来定,以R2 值越趋近于1 为好)。推荐使用专业制作曲线软件进行分析,如curve expert 1.30,根据样品O.D.值,由标准曲线查出相应的浓度,乘以稀释倍数;或用标准物的浓度与O.D.值计算出标准曲线的回归方程式,将样品的O.D.值代入方程式,计算出样品浓度,再乘以稀释倍数,即为样品的实际浓度。[典型数据]为了便于计算,尽管浓度为自变量而O.D.值为因变量,我们绘图时仍采用标准品的O.D.值作为横坐标(X 轴),标准品的浓度为纵坐标(Y 轴)。同时为了试验结果的直观性,图中提供的是原始数据而非对数值。推荐使用对数值建立标准曲线图。由于实验操作条件的不同(如操作者、移液技术、洗板技术和温度条件等),标准曲线的O.D.值会有所差异。所提供的标准曲线仅供参考,实验者需要根据自已的实验建立标准曲线。人粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒标准曲线[检测范围]31.2pg/mL-2,000pg/mL[最低检测限]12.6pg/mL此值为20 个空白样品(即标准品稀释液)测定的平均值加二倍标准差所对应的浓度。[特异性]本试剂盒用于检测GMCSF,经检测与其它相似物质无明显交叉反应。由于受到技术及样本来源的限制,不可能完成对所有相关或相似物质交叉反应检测,因此本试剂盒有可能与未经检测的其它物质有交叉反应。[回收率]分别于定值血清及血浆样本中加入一定量的GMCSF(加标样品),重复测定并计算其均值,回收率为测定值与理论值的比率。样本回收率范围(%) 平均回收率(%)血清(n=5) 87-101 95EDTA 血浆(n=5) 80-95 88肝素血浆(n=5) 90-103 97[线性]在定值血清及血浆样本内加入适量的GMCSF,并倍比稀释成1:2,1:4,1:8,1:16 的待测样本,线性范围即为稀释后样本中GMCSF 含量的测定值与理论值的比率。样本1:2 1:4 1:8 1:16血清(n=5) 85-97% 88-99% 91-101% 79-97%EDTA 血浆(n=5) 83-95% 93-102% 88-97% 90-105%肝素血浆(n=5) 90-102% 96-105% 91-99% 85-95%[精密度]精密度用样品测定值的变异系数CV 表示。CV(%) = SD/mean×100批内差:取同批次试剂盒对低、中、高值定值样本进行定量检测,每份样本连续测定20 次,分别计算不同浓度样本的平均值及SD 值。批间差:选取3 个不同批次的试剂盒分别对低、中、高值定值样本进行定量测定,每个样本使用同一试剂盒重复测定8 次,分别计算不同浓度样本的平均值及SD 值。批内差: CV批间差: CV[稳定性]经测定,试剂盒在有效期内按推荐温度保存,其活性降低率小于5%。为减小外部因素对试剂盒破坏前后检测值的影响,实验室的环境条件需尽量保持一致,尤其是实验室内温度、湿度及温育条件。其次由同一实验员来进行操作可减少人为误差。[实验流程]1、实验前标准品、试剂及样本的准备;2、加样(标准品及样本)100μL,37oC孵育2小时;3、吸弃,加检测溶液A100μL,37oC孵育1小时;4、洗板3次;5、加检测溶液B100μL,37oC孵育30分钟;6、洗板5次;7、加TMB底物90μL,37oC孵育15-25分钟;8、加终止液50μL,立即450nm读数。[说明]1、由于现有条件及科学技术水平尚不能对所有供货商提供的所有原料进行全面的鉴定与分析,本产品可能存在一定的质量技术风险。2、最终的实验结果与试剂的有效性、实验者的相关操作以及当时的实验环境密切相关,请务必准备充足的标本备份。3、不同批次的同一产品可能会有少许差别,如:检测限、灵敏度以及显色时间等,请依据试剂盒内说明书进行实验操作,网站电子版说明书仅作参考。4、只有全部使用本试剂盒配套试剂才能保证检测效果,不能混用其他制造商的产品。只有严格遵守本试剂盒的实验说明才会得到最佳的检测结果。5、在储存及温育过程中避免将试剂暴露在强光中。所有试剂瓶盖须盖紧以防止蒸发和微生物的污染,因为蛋白水解酶的干扰将导致出现错误的结果。6、刚开启的酶联板孔中可能会有少许水样物质,此为正常现象,不会对实验结果造成任何影响。请勿提前将酶标板从包装袋里拿出。7、由于操作者不熟练、操作失误或读数仪程序选用错误等有可能会导致错误结果的产生。请使用者使用该产品前仔细阅读说明书,调试好酶标仪,请使用配备有450±10nm 滤光片的酶标仪,且该酶标仪测量范围在0.001-3.000 O.D.或以上。8、同一使用者在使用同一产品时,如不同时间订购,也可能会因不同批次的少量差异产生不同结果,因此建议使用者在每次使用前均进行预实验。9、试剂盒在出厂前均经过严格检测,但由于运输条件及各实验室条件差异,可能会造成实验结果与出厂结果不一致或不同批次试剂盒批间差大的情况,对于这种情况会酌情处理。10、本试剂盒未与其他厂家同类试剂盒或不同方法检测同一目的蛋白的产品做对比,平行检测可能会存在检测结果不一致的情况。11、本操作说明同样适用于48T 试剂盒,但48T 试剂盒所有试剂减半。[警告]本试剂盒中使用了稀硫酸作为终止液,其具有轻微腐蚀性,使用时应避免接触衣物或眼、手等皮肤暴露部位。[问题解答]问题可能原因解决方案标准曲线差标准品准备不正确进行正确的标准品梯度稀释吸取及洗涤不充分充分的吸取及洗涤移液不精确检查和校正移液器精密度低洗涤不充分按说明书要求充分洗涤和浸泡混匀不充分和吸取试剂不足充分混匀和吸取试剂重复利用吸头、容器和覆膜使用加样器要更换新的吸头、使用新的容器和覆膜加样不精确检查和校正移液器O.D 值低每孔加的试剂量不精确校正移液器,精确加入试剂温育时间不正确保证充足的温育时间温育温度不正确试剂要平衡至室温并保证准确的温育温度酶标记物或底物失效通过混合酶标记物和底物,颜色应迅速显现来检查没有加入终止液按照说明书实验操作步骤加入终止液超出读数时间读数在说明书推荐的读数时间内读数样本值不正确的样本储存方式正确储存样本,使用新鲜样本进行实验不正确的样本收集和处理方法采取正确的样本收集和处理方法待测物质在样本中含量低使用新鲜样本,重复实验
标准
2015.04.27
在美国癌症研究协会(AARC)2015年年会上,来自科罗拉多大学的学者展示了最新的研究结果关于治疗结直肠癌的新潜力药物——口服水飞蓟宾(silibinin,从牛奶蓟中纯化提取),减慢结直肠癌干细胞生长的能力。水飞蓟宾治疗后采样的肿瘤干细胞重新注入到新模型上,细胞未能发展成具有同样侵袭性的肿瘤,即使在没有再服用水飞蓟宾的情况下。水飞蓟宾是存在于牛奶蓟种子内的非毒性,潜在的化学预防剂。在去年的AACR年会上Agarwal博士和他的同事发现,在细胞培养物中,水飞蓟宾影响大肠癌干细胞的形成和生存相关的细胞信号传导。现在,这个充满希望的研究扩展到这个小鼠模型上。研究人员将结直肠癌干细胞种植到喂或不喂水飞蓟宾的小鼠体内。肿瘤生长通过可见的尺寸测量,MRI扫描和肿瘤代谢率(葡萄糖的使用)来测量。随后小鼠体内的肿瘤干细胞重新注入下一代小鼠,以测量在没有水飞蓟宾影响下的肿瘤生长模式。“在水飞蓟宾喂养的小鼠肿瘤内,肿瘤干细胞数目很少,肿瘤体积较小,代谢水平较低,并表现出新的血管生长水平下降。更重要的是,水飞蓟宾喂养的小鼠的肿瘤干细胞失去了分裂繁殖的能力。”Rajesh Agarwal博士总结说道。研究人员表示他们已经在深入进行这项研究,从水飞蓟宾本身到它在结直肠癌的化学预防性。目前的研究正在验证一个重要之处:在动物模型上同时观察到可能的化学预防特性和治疗效果。具体的分子机制的发现还需要一定的研究,确认水飞蓟宾在癌症的预防和或治疗的人体临床实验也在计划之中
百态
2015.04.27
有时候癌症研究者们会考虑将来自“死亡之帽”毒蘑菇衍生的毒性α-鹅膏蕈碱作为一种潜在的癌症疗法药物,然而由于α-鹅膏蕈碱会存在引发肝脏毒性的可能,因此其作为有效的疗法往往被认为存在很多限制。近日一篇刊登在国际杂志Nature上的研究论文中,来自得克萨斯大学MD癌症研究中心的研究人员着眼于开发一种以α-鹅膏蕈碱为基础的抗体共轭药物(ADCs),他们发现ADCs可以以POLR2A基因为靶点来来有效治疗患结直肠癌的小鼠模型,这种药物会有效促进肿瘤消退以及大幅降低自身的毒性,同时ADCs还会改善对癌细胞的靶向作用,尽可能地减少对健康细胞的影响。Xiongbin Lu教授表示,当传统的肿瘤抑制基因TP53被剔除后就会引发癌症发展,而其附近的基因POLR2A同时也会被剔除;正常细胞中拥有两个拷贝的POLR2A和TP53基因,而本文研究中研究者对含有单一拷贝两个基因的癌细胞进行研究,单一拷贝的两个基因在53%的结直肠癌、62%的乳腺癌以及75%的卵巢癌中存在。POLR2A是一种包括癌细胞在内的细胞生存的必须基因,由于其仅存在单一拷贝,因此癌细胞对于该基因的抑制会异常敏感。随着POLR2A和TP53基因同时被剔除就意味着靶向作用这两个基因遗传过程的疗法变得不再那么有效,从而使得癌细胞继续旺盛成长;而揭示单一拷贝的POLR2A基因或许就可以促进癌症发展,帮助科学家们寻找新型靶点来进行癌症治疗;文章中研究者检测了ADCs的作用,发现其可以有效抑制POLR2A基因的表达,进而抑制癌症的发展。在癌症疗法中研究者们花费了很大努力来试图恢复TP53的活性,然而由于TP53信号的复杂性,目前并没有基于TP53的疗法成功转化进入临床的癌症疗法阶段。POLR2A基因可以编码一种酶类,该酶类会被α-鹅膏蕈碱所以只,研究者发现,低剂量α-鹅膏蕈碱引发的基因POLR2A的抑制会阻断癌细胞的生长,并且降低毒性。最后研究者Lu说道,我们预测,抑制基因POLR2A的表达或许可以作为一种新型的治疗手段来治疗携带常见基因突变的人类癌症
厂商
2015.04.27
wnt信号包括β catenin依赖性的经典途径和β catenin非依赖性的非经典途径,在调节动物发育和维持稳态方面具有重要作用,wnt信号失调会引起包括癌症在内的多种疾病发生。 wnt-frz之间发生相互作用能够激活非经典途径,但对于经典途径没有影响,这一过程的发生需要wnt共受体LRP5/6的作用。近日,来自同济大学的研究人员在国际学术期刊nature communication在线发表了他们关于LRP5/6 在frizzled介导的肿瘤转移中发挥抑制作用的最新研究进展。 在该项研究中,研究人员发现wnt受体frizzled及其共受体LRP5/6能够发生直接相互作用,并且这种相互作用受到LRP6胞外结构域的调节作用。同时,通过与frz直接结合,LRP5/6能够抑制frz介导的wnt非经典途径激活并进一步抑制wnt非经典途径介导的肿瘤转移过程。研究人员通过体外实验证实敲低内源性LRP5/6能够促进非侵袭性肿瘤细胞迁移,利用LRP6胞外段重组蛋白进行处理可以抑制侵袭性肿瘤细胞的体外迁移和体内转移。同时,细胞膜LRP5/6的表达水平与小鼠和人类乳腺癌转移呈负相关性。 这项研究首次揭示了膜受体frizzled与LRP5/6之间的相互作用,同时发现了LRP5/6抑制wnt非经典信号途径的分子机制,为LRP6胞外结构域在抑制肿瘤转移方面的临床应用提供了重要生物学基础
厂商
2015.04.24
肺结核是目前世界上最严重的传染病之一,每年造成9百万以上的感染以及1百万的死亡。该病的致病菌-结核杆菌具有很强的药物耐受性,长期的药物治疗很容易使得结核病成为不治之症。因此,发现新的抗结核杆菌免疫应答是治疗结核病的绝佳思路。IL-32是2005年被鉴定出来的一类细胞因子,它对于炎性因子(比如TNF-a,IL-1b)的释放具有重要的促进作用。之前的研究发现IL-32对于多种病原体具有抵抗的效果,而这一效果很可能是由于IL-32的促炎性效应。之前的研究发现:当用结核杆菌(MTB)刺激人源巨噬细胞以及外周血单核细胞时,会引起 IL-32的释放,人为抑制IL-32的表达则导致MTB的过度增长。这一结果暗示了IL-32抵抗MTB感染的作用。然而,体内的具体情况目前还不清楚。 最近,来自科罗拉多州立大学微生物系的Edward D. Chan课题组在《PNAS》杂志在线发表了他们对IL-32在MTB感染小鼠肺部过程中的作用机制的研究。 首先,作者培育了一类转基因小鼠,该小鼠在肺泡上皮细胞中特异性表达IL-32gamma。之后他们比较了野生型小鼠与转基因小鼠在MTB感染数天后的细菌定植情况。结果显示,相比于野生型小鼠,转基因小鼠肺部MTB的细菌数量明显下降,脾脏中也出现了类似的结果。之后,作者发现在长期的感染之后,转基因小鼠的存活率明显高于野生型。 由于IL-32能够通过分泌到胞外从而影响其它的细胞,作者比较了野生型与转基因小鼠肺泡巨噬细胞受MTB的影响。体外感染结果显示,相对于野生型肺泡巨噬细胞,转基因的肺泡巨噬细胞中MTB的感染水平明显下降。由于被吞噬进入胞内小体中的MTB依赖于与溶酶体融合来进行消化。作者比较了MTB进入不同类型的巨噬细胞内部后与溶酶体的共定位情况。结果显示,转基因的巨噬细胞内部出现了更高的共定位比例。这说明转基因小鼠的肺泡巨噬细胞杀伤MTB的能力大大提高了。 进一步的检测发现,在感染30天后,转基因小鼠的肺泡组织中富集了大量的TNF-a阳性的先天免疫细胞(肺泡巨噬细胞,树突状细胞等),这一比例远远高于野生型小鼠。此外,作者在转基因小鼠的肺泡组织以及外周淋巴结中发现了更多的IFN-gamma+ CD4+ 与CD8+ T细胞以及调节性T细胞。 由于IL-32具有多个mRNA剪接体,其中IL-32b具有最强的免疫抑制活性。作者向小鼠巨噬细胞系RAW264.7中转入了野生的IL-32gamma或者一个突变体,这一突变的IL-32gamma丧失了剪接形成IL-32b的能力。之后,作者分别将两种转染株进行MTB感染,结果显示:在较长时间的感染后,突变体相对于野生株其MTB感染能力更为低下。 最后,作者利用免疫组化的手段验证了在MTB感染过程中,患者肺泡组织会有大量IL-32的分泌,也验证了上述在小鼠模型上得出的结论
厂商
2015.04.24
产品类型:ELISA Kit产品名称:绵羊花生四烯酸(AA)ELISA kit英文名称:Sheep arachidonic Acid(AA) ELISA kit货号:yb-EQ027590SH规格:96T种属:Sheep其他种属:Human Mouse Rat Chicken Pig Rabbit Bovine Plant待测物名称:arachidonic Acid(AA)缩写:AA蛋白功能1:others检测范围:15.625 ng/ml-1000 ng/ml灵敏度:13.797 ng/ml反应时间:1-5h所需样本体积:50-100ul检测波长:450nm
标准
2015.04.23
产品类型:ELISA Kit产品名称:Human ATP-binding cassette sub-family A member 13(ABCA13) ELISA kit英文名称:Human ATP-binding cassette sub-family A member 13(ABCA13) ELISA kit货号:YB-EL001037HU别名:DKFZp313D2411, FLJ16398, FLJ33876, FLJ33951, ATP binding cassette transporter A13|ATP binding cassette, sub-family A (ABC1), member 13规格:96T种属:Human待测物名称:ATP-binding cassette, sub-family A (ABC1), member 13缩写:ABCA13蛋白功能1:Transport蛋白功能3:Transport检测范围:18.75 pg/ml-1200 pg/ml灵敏度:4.69 pg/ml反应时间:1-5h所需样本体积:50-100ul检测波长:450 nm
标准
2015.04.23
标记免疫疗法治疗癌症又向前迈进一大步,科学家已经证明了纳米涂层细菌能有效转运口服DNA疫苗,这种疫苗能刺激人体自身的免疫系统发挥作用并摧毁癌细胞。这是第一次纳米涂料用于经体内细菌转运口服DNA疫苗。与未经涂层的细菌相比,涂层细菌可以绕过很多“路障”,这些“路障”到目前为止限制了免疫反应,成为DNA疫苗治疗癌症面临的最大挑战。由新加坡南洋理工大学平远(音)和中国浙江大学唐谷平(音)领导的科研人员,在最近一期的《纳米通讯》上发表了相关论文。通常来说,免疫疗法被认为是目前多种治疗癌症方法中的一种潜在替代化疗和放疗的可行方法,化疗和放疗能直接攻击并摧毁癌细胞,但也在治疗过程中损害了正常细胞。因为免疫疗法激发身体自身的免疫系统来对准并消灭癌细胞,因此,比其他疗法更加安全,带来的副作用也更少。“我们工作的最重要贡献,是为有效增加口服癌症疫苗生物利用程度提供了一个重要的运输方案。”平远说。研究人员正在攻克的一种DNA疫苗被称为NP/SAL,能抑制肿瘤血管的生成。许多肿瘤分泌的血管生长因子如血管内皮生长因子(VEGF)来促进血管生成,最终导致肿瘤转移。NP/SAL疫苗能刺激免疫系统产生T细胞(白细胞)和细胞因子(化学信使),反过来能干扰VEGF通路,进而减少血管形成并最终抑制肿瘤生长。前提是将疫苗运送到合适位置,将疫苗接种到沙门氏菌中后,细菌就以典型的方式侵入人体,人体感染细菌、细菌在体内繁殖并传播它们的DNA从而达到免疫效果。在疫苗能记起免疫反应之前,必须克服两个主要障碍——巨噬细胞的吞噬,以及胃与小肠的高酸性环境,只有一小部分原始菌种能通过两道屏障,这也是到目前为止疫苗失败的主要原因。新论文指出,研究人员第一次证明了纳米涂层细菌比未涂层细菌更容易攻克这两道屏障,也更容易激发更强的免疫反应。研究人员发现,在60%被注射疫苗的小鼠中,存活了35天而没有肿瘤扩散,小鼠几乎没有体重减少,反应了这种疫苗的低毒性。研究人员希望纳米涂层细菌DNA疫苗载体策略能被应用于研发治疗各种癌症的疫苗。“我们希望这种疫苗能在未来3—5年内以传统小瓶的药剂形式应用于临床。”平远说,“除了沙门氏菌,还有不少细菌可供选择,我们希望设计出不同类型的疫苗战略,个性化纳米药物治疗免疫性疾病将迎来新的曙光
百态
2015.04.22
一项新的研究发现,胰腺癌细胞可以通过过表达一个名为E47的蛋白,从而被诱导转变回正常的细胞。E47可以与特定的DNA序列结合,来控制掌管生长和分化的一些基因。这项研究为美国每年超过4万死于胰腺癌的患者们提供了拥有新治疗方法的希望。"这是第一次,我们发现过表达一个基因,就可以降低胰腺腺癌细胞促进肿瘤发展的潜在性,通过将这些癌症细胞重新编程回它们原始的细胞类型。因此,胰腺细胞保留有基因记忆,我们希望可以开发利用。"Sanford-Burnham发育,衰老和再生项目的兼职教授Pamela Itkin-Ansari说,她也是这项研究的主要作者。此研究在于4月20日发表在Pancreas杂志上。E47将时钟拨了回去这项研究是几个大学联合合作的成果,包括Sanford-Burnham,圣迭戈大学(Itkin-Ansari负责联合任命)和普渡大学,研究人员们造出了人胰腺导管腺癌细胞系,且E47水平高于正常水平。升高的E47导致了细胞停滞在G0/G1生长期,分化回了腺泡细胞表型。体内实验显示,当这些重新编程的癌症细胞被打入老鼠体内,这些细胞形成肿瘤的能力,与未处理的胰腺癌细胞相比,大大减低。"目前,胰腺腺癌的治疗方法是一些细胞毒性药物,患者在确证后的平均存活时间仅为6个月,治疗的改进时间以天来计算,"圣迭戈大学莫尔斯癌症中心外科教授,美国国家癌症研究所胰腺癌专责小组联合主席Andrew M. Lowy说。"我们可以将癌症细胞去分化为没有任何危害的细胞表型,这项发现十分地鼓舞人心。事实上,先前已经有细胞分化的治疗方法,成功地用于治疗急性早幼粒细胞白血病(APL)和某些神经细胞瘤。""我们的下一步将测试病人的肿瘤样本,看E47是否可以引起相似的结果,从而为抗击这种高致死率的疾病提供新的治疗方法。"Itkin-Ansari说。"此外,我们也将筛选可以引起E47过表达的分子-蛋白药物。"胰腺腺癌胰腺腺癌是最常见的一种胰腺癌症。这种癌症主要由癌基因Kras突变引起,从而导致了消化酶分泌细胞(腺泡细胞)分化为一种不稳定的导管样细胞类型,这种细胞有致癌性。胰腺腺癌常常被称为"沉默"癌症,因为它极少表现出任何症状,当它引起体重下降,腹痛和黄疸时,往往已经是癌症晚期了
厂商
2015.04.22
什么是夹心ELISA?答:夹层ELISA采用对样本中分析物的特异的抗体作为固定化的捕获抗体,然后用第二个带标记的抗体作检测,检测抗体对分析物也是专一的。当参照标准曲线时,试验数据值是定量的。
标准
2015.04.21
什么是直接ELISA?答:直接ELISA是将被测的分析物直接包膜在微孔板表面,然后用测试抗体来证实被测分析物的存在。当与重组蛋白(标准曲线)进行参照时,试验数据值是半定量的。
标准
2015.04.21
近日,一项刊登在国际杂志Neuron上的研究论文中,来自日本东京工业大学等处的科学家们通过研究发现,当眼睛中的神经元长时间暴露于光下后,其会改变特殊分子的水平,随后研究者又鉴别出了一种特殊的反馈信号机制或许是引发这一改变的原因,因此研究者或可利用先天性的神经元特性来保护眼部神经元免于退化或细胞死亡。神经元间突触的改变会促进机体更加适应环境的改变,然而截止到现在研究者尚不清楚隐藏在“突触可塑性”下的信号机制;本文中研究者就揭示了突触可塑性发生的详细分子机制。Atsushi Sugie教授表示,我们所鉴别出的突触改变可以反映出先天性的神经元特性,而这些特性往往会保护神经元免于过度的刺激;通过增强这些神经元特性研究者就可以保护神经元免于退化和细胞死亡。近来有研究表明,果蝇突触前膜区域的改变,即活性区的改变会控制突触的功能;研究者将果蝇暴露于不同的光下,随后比较其光感受器活性区域的差异;位于突触前膜的T形结构可以“拴着”突触小泡,同时控制神经递质向突触后神经元的传递释放,通过对T形结构关键的蛋白进行标记研究者发现,当其它因素未改变的时候,一系列的活性区域蛋白水平却下降了,随后研究者还发现了相应的结构蛋白的缺失以及T形结构数量的减少。最后研究者还表示,反馈信号机制主要负责上述改变,而且其依赖于信号蛋白Wnt,相关研究为理解大脑功能,即学习和记忆的发生背后的分子机制提供了一定的思路;后期研究者将去调查如何通过修饰Wnt信号来操控突触的可塑性,同时这也为开发治疗神经变性疾病或精神疾病的新型疗法提供了帮助
厂商
2015.04.20
先兆子痫是一种影响5%-8%的美国孕妇的疾病,该疾病引发的并发症通常会导致女性怀孕早期紧急剖腹产,这样才能挽救婴儿和母亲的生命,而科学家们认为先兆子痫是由一系列因素所引发的,包括浅胎盘等,浅胎盘和婴儿母源性血管不足直接相关。近日一篇发表在国际杂志PNAS上的研究论文中,来自密苏里大学的研究者通过研究成功培育生长了胎盘细胞,其可以帮助更好地进行先兆子痫发病机制的研究。研究者Michael Roberts表示,胎盘细胞是一种未知形式的人类胚胎干细胞,这种新型干细胞可以帮助研究先兆子痫的发病机制及人类生育过程的其它过程。研究者将这种干细胞称之为骨形成蛋白干细胞,相比正常的胚胎干细胞而言其非常强劲而且容易对其进行操作;骨形成蛋白干细胞是一种介于胚胎干细胞和最后发育形成后的细胞间的一种过渡状态的细胞,研究者可以利用这种干细胞来理解胚胎的形成过程以及进行诸如先兆子痫等出生前疾病发病机制的研究。胚胎干细胞具有多能性,这就意味着研究者可以利用其分化形成一系列不同的细胞,比如肌肉细胞、骨细胞等,这项研究中研究者想利用短期添加骨形成蛋白-4使胚胎干细胞分化生长为胚胎细胞,同时研究者还添加了两种药物,其可以暂时抑制干细胞多能状态的关键生化反应。如果不形成胎盘细胞,干细胞就会生长成为以前的一种未被观察的状态,也就是研究者所谓的骨形成蛋白干细胞,相比传统干细胞而言这种干细胞很容易在实验室进行操作,而且容易生长,具有较强的均一性,这就意味着培养中的细胞都会表现出一致的遗传信息。研究者Roberts表示,此前传统观念认为胚胎干细胞的过渡会径直从干细胞到最终的分化细胞,而本文中新型干细胞的发现则可以帮助研究者认识到胚胎干细胞是以许多不同过渡阶段的细胞而存在的,相关研究就可以帮助研究人员未来进行更加有效的干细胞研究以及开发治疗婴儿出生前相关疾病的新型靶向疗法
百态
2015.04.20
近日,发表于国际杂志JAMA Psychiatry上的研究论文中,来自华盛顿大学医学院的研究者通过研究表示,抑郁症和2型糖尿病或许都和痴呆症的风险增加直接相关,而该风险在那些被诊断既是抑郁症又是糖尿病的患者中尤其高。糖尿病和严重抑郁在西方国家非常常见,大约有20%的糖尿病患者会患上抑郁症。研究者Dimitry Davydow表示,我们对240万丹麦居民进行了相关研究,检测了抑郁症患者、2型糖尿病患者以及同时患两种病的患者相比未患病的患者患痴呆症的风险,参与者年龄都在50岁及以上,其在2007年至2013年均无痴呆症病症表现。总的来讲,研究小组中有19.4%的个体为抑郁症(477,133名个体),有9.1%的个体为2型糖尿病(223,174名个体),有3.9%的个体均患上了两种病(95,691名个体),起初诊断为2型糖尿病的个体年龄为63.1岁,最初被诊断为抑郁症的年龄则为58.5岁。研究者表示,在研究期间有2.4%的个体患上了痴呆症(59,663名个体),平均年龄为81岁,在患痴呆症的个体中,15729名个体(26.4%)仅为抑郁症患者,6466名个体(10.8%)仅为2型糖尿病患者,同时4022名个体(6.7%)均患上了两种疾病。研究结果表明,仅患2型糖尿病和痴呆症风险增加20%直接相关,而仅患抑郁症则和痴呆症风险增加83%相关,共患两种病则和痴呆症风险增加117%相关,而且痴呆症的患病风险似乎在65岁以上的参与者中尤其高。从慢性病增加社会负担的角度考虑,未来研究中研究者需要阐明和抑郁症,2型糖尿病发生相关的病理学机制,以及揭示引发患者不良后果的原因,比如痴呆症等,从而为开发新型干预措施来抑制相关疾病并发症的发生提供新的线索和思路
百态
2015.04.17
近日,一项刊登在国际杂志ecancermedicalscience上的研究论文中,来自ReDO(对药物进行重新定向的肿瘤研究计划)计划小组的研究者通过研究表示,一种常见的抗真菌疗法或可用于治疗癌症。ReDO计划即国际抗癌科学家联合在一起进行的一项大型研究,致力于开发利用常见药物来作为新型的癌症疗法。该项计划如今已经发表了关于药物开发的数篇研究报道,而且这些药物具有一定的临床证据足以进行临床试验。其中伊曲康唑就是一种用于治疗广谱真菌感染的药物,包括皮肤和指甲的感染等,该药物同时或可作为一种新型的抗癌疗法。Pan Pantziarka指出,伊曲康唑在治疗一系列癌症患者上表现出了巨大潜力,尤其是非小细胞肺癌及某些罕见的恶性肿瘤。伊曲康唑是一种非专利制剂,其非常廉价而且有证据表明其可以有效抑制前列腺癌的转移,研究者表示,这或许就可以使得伊曲康唑成为极具吸引力的新型癌症药物,不仅对于中低收入国家非常重要,而且对于卫生系统不堪重负的国家也是一项福利。但在抗真菌药物进入抗癌市场前却难免会遇到一些障碍,目前研究者正在收集对药物重新定向的数据,旨在得到医学界的关注。这些重新定向的抗癌药物,比如抗真菌药物或止痛药或许都可以应用于未来的癌症药物开发研究过程,研究者希望提高科学家对重新定向药物的认知来使得更多抗癌药物被发现
厂商
2015.04.17