大鼠ELISA试剂盒
大鼠烟酰胺腺嘌呤二核苷酸磷酸(NADPH)酶联免疫吸附测定试剂盒
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货号
FY-ER5205样本类型
Serum, Plasma, Tissue homogenate and Other biological samples检测时间
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产品详情
英文名称:
Rat NADPH(Nicotinamide Adenine Dinucleotide Phosphate) ELISA Kit
种属:
Rat
别称:
NADPH
检测方法:
Competitive
标准品:
2000ng/mL
灵敏度:
18.75 ng/mL
检测范围:
31.25-2000ng/mL
反应时间:
1.5H
样品类型:
Serum, Plasma, Tissue homogenate and Other biological samples
研究领域:
Enzyme & Kinase;Apoptosis;
测试原理:
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with NADPH protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to NADPH. Next,Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of NADPH in the samples is then determined by comparing the OD of the samples to the standard curve.
技术数据
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration | OD1 | OD2 | Average OD | Corrected |
|---|---|---|---|---|
| 2000 | 0.19 | 0.192 | 0.191 | |
| 1000 | 0.326 | 0.329 | 0.328 | |
| 500 | 0.541 | 0.546 | 0.544 | |
| 250 | 0.875 | 0.882 | 0.879 | |
| 125 | 1.303 | 1.303 | 1.303 | |
| 62.5 | 1.695 | 1.701 | 1.698 | |
| 31.25 | 2.071 | 2.079 | 2.075 | |
| 0 | 2.511 | 2.53 | 2.521 |
线性
Samples were spiked with high concentrations of target proteins and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 95-102 | 88-97 | 93-107 |
| Average (%) | 102 | 89 | 98 | |
| 1:4 | Range (%) | 83-96 | 88-97 | 85-94 |
| Average (%) | 87 | 95 | 93 | |
| 1:8 | Range (%) | 97-105 | 96-105 | 87-96 |
| Average (%) | 100 | 103 | 94 | |
| 1:16 | Range (%) | 85-97 | 81-96 | 92-101 |
| Average (%) | 88 | 88 | 94 |
精密度
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.
| Item | Intra-assay Precision | Intra-assay Precision |
|---|---|---|
| Sample | 3 | 3 |
| Replicate | 9 | 18 |
| CV(%) | 5 | 8 |
回收率
The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range(%) | Average Recovery(%) |
|---|---|---|
| Serum (n=8) | 80-96 | 88 |
| EDTA plasma (n=8) | 82-98 | 90 |
| Cell culture media (n=8) | 84-98 | 91 |
检测流程图
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