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牛转录因子P65(RELA) ELISA kit货号:SY-EL019553BO
点击次数：438发布时间：2013/3/14 17:10:49
更新日期：2013/11/5 9:36:21
所 在 地：中国大陆
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详细内容
MGC131774, NFKB3, p65, nuclear factor of kappa light polypeptide gene enhancer in B-cells 3|v-rel avian reticuloendotheliosis viral oncogene homolog A （nuclear factor of kappa light polypeptide gene 货号： CSB-EL019553BO 中文价格： 4200 规格： 96T 种属： Bovine 待测物名称： v-rel reticuloendotheliosis viral oncogene homolog A （avian） 缩写： RELA 蛋白功能1： Transcription/Transcription regulation 蛋白功能3： Transcription 检测范围： Request Information 灵敏度： Request Information 反应时间： 1-5h 所需样本体积： 50-100ul 检测波长： 450nm 下载： 注：产品说明书可能有改进，请直接与CUSABIO产品专员联系，索取*新版说明书！
用途： For research use only. Not for diagnostic use. 原理： This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for RELA has been pre-coated onto a microplate. Standards ａnd samples are pipetted into the wells ａnd any R......＋This assay employs the quantitative sandwich enzyme immunoassay technique. Antibody specific for RELA has been pre-coated onto a microplate. Standards ａnd samples are pipetted into the wells ａnd any RELA present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for RELA 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 RELA 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 Bovine RELA. No significant cross-reactivity or interference between Bovine RELA 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 immediat......＋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......＋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......＋
