产品名称: 小鼠腺苷高半胱氨酸酶(AHCY)ELISA kit
英文名称: Mouse Adenosylhomocysteinase,AHCY ELISA kit
别名: SAHH, S-adenosylhomocysteine hydrolase
货号: CSB-EL001474MO
规格: 96T
种属: Mouse
待测物名称: S-adenosylhomocysteine hydrolase
缩写: AHCY
蛋白功能1: Biosynthesis/Metabolism
蛋白功能2: Amino-acid biosynthesis and metabolism
蛋白功能3: One-carbon metabolism
样本类型: serum, plasma, tissue homogenates, cell lysates
检测范围: 28.12 pg/ml-1800 pg/ml
灵敏度: 7.03 pg/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 AHCY has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any AHCY present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for AHCY 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 AHCY 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 Mouse AHCY. No significant cross-reactivity or interference between Mouse AHCY 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 AHCY 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:
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抗体
产品名称 货号 规格
Rabbit anti-human S-adenosylhomocysteine hydrolase polyclonal Antibody CSB-PA001474GA01HU 150ul
蛋白
产品名称 货号 规格 蛋白来源
Recombinant Pseudomonas putida Adenosylhomocysteinase(ahcY) CSB-EP001474FGA 1mg E.coli
Recombinant Pseudomonas putida Adenosylhomocysteinase(ahcY) CSB-YP001474FGA 1mg Yeast
Recombinant Xylella fastidiosa Adenosylhomocysteinase(ahcY) CSB-EP001474XBM 1mg E.coli
Recombinant Xylella fastidiosa Adenosylhomocysteinase(ahcY) CSB-YP001474XBM 1mg Yeast
Recombinant Methylobacterium sp. Adenosylhomocysteinase(ahcY) CSB-EP001474MTE 1mg E.coli
Recombinant Methylobacterium sp. Adenosylhomocysteinase(ahcY) CSB-YP001474MTE 1mg Yeast
Recombinant Halobacterium salinarum Adenosylhomocysteinase(ahcY) CSB-EP001474HTL 1mg E.coli
Recombinant Halobacterium salinarum Adenosylhomocysteinase(ahcY) CSB-YP001474HTL 1mg Yeast
Recombinant Pseudomonas putida Adenosylhomocysteinase(ahcY) CSB-EP001474FGC 1mg E.coli
Recombinant Pseudomonas putida Adenosylhomocysteinase(ahcY) CSB-YP001474FGC 1mg Yeast
Recombinant Brucella suis Adenosylhomocysteinase(ahcY) CSB-EP001474BWU 1mg E.coli
Recombinant Brucella suis Adenosylhomocysteinase(ahcY) CSB-YP001474BWU 1mg Yeast
Recombinant Streptomyces griseus subsp. griseus Adenosylhomocysteinase(ahcY) CSB-EP001474FOF 1mg E.coli
Recombinant Streptomyces griseus subsp. griseus Adenosylhomocysteinase(ahcY) CSB-YP001474FOF 1mg Yeast
Recombinant Aquifex aeolicus Adenosylhomocysteinase(ahcY) CSB-EP001474DNV 1mg E.coli
Recombinant Aquifex aeolicus Adenosylhomocysteinase(ahcY) CSB-YP001474DNV 1mg Yeast
Recombinant Anopheles gambiae Adenosylhomocysteinase(Ahcy13) CSB-EP001474BZL 1mg E.coli
Recombinant Anopheles gambiae Adenosylhomocysteinase(Ahcy13) CSB-YP001474BZL 1mg Yeast
Recombinant Pyrococcus horikoshii Adenosylhomocysteinase(ahcY) CSB-EP001474FHX 1mg E.coli
Recombinant Pyrococcus horikoshii Adenosylhomocysteinase(ahcY) CSB-YP001474FHX 1mg Yeast
Recombinant Thermotoga maritima Adenosylhomocysteinase(ahcY) CSB-EP001474TNJ 1mg E.coli
Recombinant Thermotoga maritima Adenosylhomocysteinase(ahcY) CSB-YP001474TNJ 1mg Yeast
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