产品名称: 人凋亡抑制因子4(API4)ELISA Kit
英文名称: Human apoptosis inhibitor 4 (API4) ELISA kit
别名: API4, EPR-1, apoptosis inhibitor 4|baculoviral IAP repeat-containing protein 5|survivin variant 3 alpha
货号: CSB-EL002706HU
规格: 96T
种属: Human
待测物名称: baculoviral IAP repeat-containing 5
缩写: BIRC5
蛋白功能1: Apoptosis/Autophagy
蛋白功能3: Apoptosis
样本类型: serum, plasma, tissue homogenates
检测范围: 12.5 pg/ml-800 pg/ml
灵敏度: 3.13 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 BIRC5 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any BIRC5 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for BIRC5 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 BIRC5 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 BIRC5. No significant cross-reactivity or interference between Human BIRC5 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 BIRC5 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
Rat Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit Rat Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit CSB-EL002706RA 96T
Pig Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit Pig Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit CSB-EL002706PI 96T
Mouse Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit Mouse Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit CSB-EL002706MO 96T
Dog Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit Dog Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit CSB-EL002706DO 96T
Cat Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit Cat Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit CSB-EL002706CA 96T
Bovine Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit Bovine Baculoviral IAP repeat-containing protein 5(BIRC5) ELISA kit CSB-EL002706BO 96T
抗体
产品名称 货号 规格
Rabbit anti-human baculoviral IAP repeat-containing 5 polyclonal Antibody CSB-PA002706GA01HU 150ul
蛋白
产品名称 货号 规格 蛋白来源
Recombinant Mouse Baculoviral IAP repeat-containing protein 5(Birc5) CSB-EP002706MO 1mg E.coli
Recombinant Mouse Baculoviral IAP repeat-containing protein 5(Birc5) CSB-YP002706MO 1mg Yeast
Recombinant Human Baculoviral IAP repeat-containing protein 5(BIRC5) CSB-EP002706HU 1mg E.coli
Recombinant Human Baculoviral IAP repeat-containing protein 5(BIRC5) CSB-YP002706HU 1mg Yeast