小鼠ELISA试剂盒
小鼠磷酸化核因子κB抑制蛋白α(pIκBα)酶联免疫吸附测定试剂盒
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货号
FY-EM3153样本类型
Tissue homogenate and Cell lysate检测时间
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产品详情
英文名称:
Mouse pIκBα(Phospho-Inhibitory Subunit of NF Kappa B Alpha) ELISA Kit
种属:
Mouse
别称:
pIκBα
检测方法:
Sandwich
标准品:
10ng/mL
灵敏度:
0.1 ng/mL
检测范围:
0.16-10ng/mL
反应时间:
3.5H
样品类型:
Tissue homogenate and Cell lysate
研究领域:
Cell Biology
测试原理:
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to pIκBα. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to pIκBα. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain pIκBα, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. 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 pIκBα 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 |
|---|---|---|---|---|
| 10 | 2.394 | 2.409 | 2.402 | 2.358 |
| 5 | 1.709 | 1.716 | 1.713 | 1.673 |
| 2.5 | 0.993 | 1.001 | 0.997 | 0.957 |
| 1.25 | 0.486 | 0.486 | 0.486 | 0.45 |
| 0.63 | 0.247 | 0.247 | 0.247 | 0.211 |
| 0.32 | 0.124 | 0.124 | 0.124 | 0.088 |
| 0.16 | 0.077 | 0.077 | 0.077 | 0.041 |
| 0 | 0.036 | 0.036 | 0.036 | 0 |
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 (%) | 80-96 | 85-101 | 85-92 |
| Average (%) | 86 | 97 | 88 | |
| 1:4 | Range (%) | 89-102 | 80-96 | 82-96 |
| Average (%) | 92 | 89 | 85 | |
| 1:8 | Range (%) | 85-97 | 87-99 | 87-103 |
| Average (%) | 85 | 92 | 89 | |
| 1:16 | Range (%) | 80-97 | 96-107 | 95-102 |
| Average (%) | 81 | 97 | 100 |
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) | 80-95 | 87 |
| EDTA plasma (n=8) | 81-95 | 88 |
| Cell culture media (n=8) | 87-99 | 93 |
检测流程图
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