人血红蛋白α亚基(HB-α)ELISA Kit

 产品名称： 人血红蛋白α亚基(HB-α)ELISA Kit
 
 英文名称： Human α-subunits of hemoglobin,HB-α ELISA Kit
 
 货号： CSB-E13105h
 
 规格： 96T
 
 种属： Human
 
 待测物名称： hemoglobin, alpha 2
 
 缩写： HBA2
 
 蛋白功能1： Transport
 
 蛋白功能2： Anion transport
 
 蛋白功能3： Oxygen transport
 
 检测范围： 10.94 ng/ml-700ng/ml
 
 灵敏度： 5.47ng/ml
 
 反应时间： 1-5h
 
 所需样本体积： 50-100ul
 
 检测波长： 450 nm
 
 用途： For research use only. Not for diagnostic use.
 
 原理 ： This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for HBA2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HBA2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for HBA2 is added to the wells. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of HBA2 bound in the initial step. The color development is stopped and the intensity of the color is measured. 
 特异性： This assay has high sensitivity and excellent specificity for detection of Human HBA2. No significant cross-reactivity or interference between Human HBA2 and analogues was observed. 
 精密度： Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
 
 样本搜集及储存： Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 15 minutes at 1000 ×g. Remove serum and assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 ×g at 2-8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
 
 检测步骤： Bring all reagents and samples to room temperature before use. Centrifuge the sample again after thawing before the assay. It is recommended that all samples and standards be assayed in duplicate.
1. Prepare all reagents, working standards, and samples as directed in the previous sections.
2. Refer to the Assay Layout Sheet to determine the number of wells to be used and put any remaining wells and the desiccant back into the pouch and seal the ziploc, store unused wells at 4°C.
3. Add 100μl of standard and sample per well. Cover with the adhesive strip provided. Incubate for 2 hours at 37°C. A plate layout is provided to record standards and samples assayed.
4. Remove the liquid of each well, don't wash.
5. Add 100μl of Biotin-antibody (1x) to each well. Cover with a new adhesive strip. Incubate for 1 hour at 37°C. (Biotin-antibody (1x) may appear cloudy. Warm up to room temperature and mix gently until solution appears uniform.)
6. Aspirate each well and wash, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (200μl) using a squirt bottle, multi-channel pipette, manifold dispenser, or autowasher, and let it stand for 2 minutes, complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining wash Buffer by aspirating ordecanting. Invert the plate and blot it against clean paper towels.
7. Add 100μl of HRP-avidin (1x) to each well. Cover the microtiter plate with a new adhesive strip. Incubate for 1 hour at 37°C.
8. Repeat the aspiration/wash process for five times as in step 6.
9. Add 90μl of TMB Substrate to each well. Incubate for 15-30 minutes at 37°C. Protect from light.
10. Add 50μl of Stop Solution to each well, gently tap the plate to ensure thorough mixing.
11. Determine the optical density of each well within 5 minutes, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. Subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
 
 结果计算： Using the professional soft "Curve Expert 1.3" to make a standard curve is recommended, which can be downloaded from our web.
Average the duplicate readings for each standard and sample and subtract the average zero standard optical density.
Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the HBA2 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data.
If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
 
相关产品：
 
产品名称 英文名称 货号 规格
 Goat Hemoglobin subunit alpha-2(HBA2) ELISA kit  Goat Hemoglobin subunit alpha-2(HBA2) ELISA kit  CSB-EL010148GO  96T 
        
蛋白 
产品名称 货号 规格 蛋白来源
 Recombinant Gadus morhua Hemoglobin subunit alpha-2(hba2)  CSB-EP010148GER  1mg  E.coli 
        
 Recombinant Gadus morhua Hemoglobin subunit alpha-2(hba2)  CSB-YP010148GER  1mg  Yeast 
        
 Recombinant Tachyglossus aculeatus aculeatus Hemoglobin subunit alpha-2  CSB-EP010148TKU  1mg  E.coli 
        
 Recombinant Tachyglossus aculeatus aculeatus Hemoglobin subunit alpha-2  CSB-YP010148TKU  1mg  Yeast 
        
 Recombinant Xenopus borealis Hemoglobin subunit alpha-2(hba2)  CSB-EP010148XAO  1mg  E.coli 
        
 Recombinant Xenopus borealis Hemoglobin subunit alpha-2(hba2)  CSB-YP010148XAO  1mg  Yeast 
        
 Recombinant Pleurodeles waltl Hemoglobin subunit alpha-2  CSB-EP010148EWZ  1mg  E.coli 
        
 Recombinant Pleurodeles waltl Hemoglobin subunit alpha-2  CSB-YP010148EWZ  1mg  Yeast 
        
 Recombinant Xenopus laevis Hemoglobin subunit alpha-2(hba2)  CSB-EP010148XBE  1mg  E.coli 
        
 Recombinant Xenopus laevis Hemoglobin subunit alpha-2(hba2)  CSB-YP010148XBE  1mg  Yeast 
        
 Recombinant Bos mutus grunniens Hemoglobin subunit alpha-2  CSB-EP010148BPD  1mg  E.coli 
        
 Recombinant Bos mutus grunniens Hemoglobin subunit alpha-2  CSB-YP010148BPD  1mg  Yeast 
        
 Recombinant Notothenia angustata Hemoglobin subunit alpha-2(hba2)  CSB-EP010148NEP  1mg  E.coli 
        
 Recombinant Notothenia angustata Hemoglobin subunit alpha-2(hba2)  CSB-YP010148NEP  1mg  Yeast 
        
 Recombinant Notothenia coriiceps neglecta Hemoglobin subunit alpha-2(hba2)  CSB-EP010148NHX  1mg  E.coli 
        
 Recombinant Notothenia coriiceps neglecta Hemoglobin subunit alpha-2(hba2)  CSB-YP010148NHX  1mg  Yeast 
        
 Recombinant Arctocephalus galapagoensis Hemoglobin subunit alpha-2  CSB-EP010148ANR  1mg  E.coli 
        
 Recombinant Arctocephalus galapagoensis Hemoglobin subunit alpha-2  CSB-YP010148ANR  1mg  Yeast 
        
 Recombinant Trematomus newnesi Hemoglobin subunit alpha-2(hba2)  CSB-EP010148TPO  1mg  E.coli 
        
 Recombinant Trematomus newnesi Hemoglobin subunit alpha-2(hba2)  CSB-YP010148TPO  1mg  Yeast 
        
 Recombinant Varanus exanthematicus albigularis Hemoglobin subunit alpha-2  CSB-EP010148VAM  1mg  E.coli 
        
 Recombinant Varanus exanthematicus albigularis Hemoglobin subunit alpha-2  CSB-YP010148VAM  1mg  Yeast 
        
 Recombinant Uromastyx hardwickii Hemoglobin subunit alpha-2  CSB-EP010148UAV  1mg  E.coli 
        
 Recombinant Uromastyx hardwickii Hemoglobin subunit alpha-2  CSB-YP010148UAV  1mg  Yeast 
        
 Recombinant Torpedo marmorata Hemoglobin subunit alpha-2  CSB-EP010148TGV  1mg  E.coli 
        
 Recombinant Torpedo marmorata Hemoglobin subunit alpha-2  CSB-YP010148TGV  1mg  Yeast 
        
 Recombinant Varecia variegata Hemoglobin subunit alpha-2  CSB-EP010148VAN  1mg  E.coli 
        
 Recombinant Varecia variegata Hemoglobin subunit alpha-2  CSB-YP010148VAN  1mg  Yeast 
        
 Recombinant Triturus cristatus Hemoglobin subunit alpha-2  CSB-EP010148TIS  1mg  E.coli 
        
 Recombinant Triturus cristatus Hemoglobin subunit alpha-2  CSB-YP010148TIS  1mg  Yeast 
        
 Recombinant Catharacta maccormicki Hemoglobin subunit alpha-2  CSB-EP010148CER  1mg  E.coli 
        
 Recombinant Catharacta maccormicki Hemoglobin subunit alpha-2  CSB-YP010148CER  1mg  Yeast 
        
 Recombinant Cottoperca gobio Hemoglobin subunit alpha-2(hba2)  CSB-EP010148CQZ  1mg  E.coli 
        
 Recombinant Cottoperca gobio Hemoglobin subunit alpha-2(hba2)  CSB-YP010148CQZ  1mg  Yeast 
        
 Recombinant Telmatobius peruvianus Hemoglobin subunit alpha-2  CSB-EP010148TBE  1mg  E.coli 
        
 Recombinant Telmatobius peruvianus Hemoglobin subunit alpha-2  CSB-YP010148TBE  1mg  Yeast 
        
 Recombinant Anarhichas minor Hemoglobin subunit alpha-2(hba2)  CSB-EP010148AJT  1mg  E.coli 
        
 Recombinant Anarhichas minor Hemoglobin subunit alpha-2(hba2)  CSB-YP010148AJT  1mg  Yeast 
        
 Recombinant Gobionotothen gibberifrons Hemoglobin subunit alpha-2(hba2)  CSB-EP010148GGX  1mg  E.coli 
        
 Recombinant Gobionotothen gibberifrons Hemoglobin subunit alpha-2(hba2)  CSB-YP010148GGX  1mg  Yeast 
        
 Recombinant Goat Hemoglobin subunit alpha-2(HBA2)  CSB-EP010148GO  1mg  E.coli 
        
 Recombinant Goat Hemoglobin subunit alpha-2(HBA2)  CSB-YP010148GO  1mg  Yeast