快速检测系列ELISA试剂盒
一步法人血管活性肠肽(VIP)酶联免疫吸附测定试剂盒
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货号
FY-EH2264S样本类型
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=50μL检测时间
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产品详情
英文名称:
EasyPro Human VIP(Vasoactive Intestinal Peptide) ELISA Kit
种属:
Human
别称:
PHM27
Uniport ID:
P01282
检测方法:
Competitive
标准品:
500pg/mL
灵敏度:
4.69 pg/mL
检测范围:
7.82-500pg/mL
反应时间:
45min
样品类型:
Serum, Plasma, Tissue homogenate and Other biological samples；Sample Volume=50μL
研究领域:
Nutrition metabolism
测试原理:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with VIP Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for VIP are added to the micro ELISA plate wells. VIP in samples (or standards) competes with a fixed amount of VIP on the solid phase supporter for sites on the HRP linked detection antibody specific to VIP. Excess conjugate and unbound sample or standard are washed from the plate. The substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450±2 nm. The concentration of VIP 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 |
|---|---|---|---|---|
| 500 | 0.162 | 0.164 | 0.163 | |
| 250 | 0.3 | 0.304 | 0.302 | |
| 125 | 0.519 | 0.529 | 0.524 | |
| 62.5 | 0.85 | 0.864 | 0.857 | |
| 31.25 | 1.271 | 1.276 | 1.274 | |
| 15.63 | 1.728 | 1.753 | 1.741 | |
| 7.82 | 2.037 | 2.058 | 2.048 | |
| 0 | 2.517 | 2.522 | 2.52 |
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 (%) | 93-102 | 90-103 | 85-92 |
| Average (%) | 94 | 103 | 88 | |
| 1:4 | Range (%) | 86-92 | 97-106 | 82-96 |
| Average (%) | 90 | 99 | 88 | |
| 1:8 | Range (%) | 96-105 | 82-90 | 87-105 |
| Average (%) | 105 | 84 | 92 | |
| 1:16 | Range (%) | 89-97 | 92-101 | 87-92 |
| Average (%) | 93 | 96 | 88 |
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) | 87-97 | 90 |
| EDTA plasma (n=8) | 81-95 | 88 |
| Cell culture media (n=8) | 83-95 | 89 |
检测流程图
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