EasyPro Series ELISA Kits
EasyPro Zebrafish TNFa(Tumor Necrosis Factor Alpha) ELISA Kit
Product No
FY-EZe1060SSample Type
serum, plasma, tissue homogenates, cell culture supernates and other biological fluidsAssay Time
45minPurchase details
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Product Details
Product Name in English:
EasyPro Zebrafish TNFa(Tumor Necrosis Factor Alpha) ELISA Kit
Species:
Zebrafish
Assay Type:
Sandwich
Standard:
200 pg/mL
Sensitivity:
1.23 pg/mL
Detection Range:
3.13-200 pg/mL
Assay Time:
45min
Sample Type:
serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Test Principle:
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with TNFa Samples (or Standards) and Horseradish Peroxidase (HRP) linked antibody specific for TNFa are added to the micro ELISA plate wells. TNFa in samples (or standards) competes with a fixed amount of TNFa on the solid phase supporter for sites on the HRP linked detection antibody specific to TNFa. 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 TNFa in the samples is then determined by comparing the OD of the samples to the standard curve.
Technical Data
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 |
|---|---|---|---|---|
| 200 | 2.313 | 2.333 | 2.323 | 2.294 |
| 100 | 1.519 | 1.542 | 1.531 | 1.502 |
| 50 | 0.888 | 0.89 | 0.889 | 0.86 |
| 25 | 0.405 | 0.408 | 0.407 | 0.378 |
| 12.5 | 0.226 | 0.228 | 0.227 | 0.198 |
| 6.25 | 0.126 | 0.127 | 0.127 | 0.098 |
| 3.13 | 0.079 | 0.079 | 0.079 | 0.05 |
| 0 | 0.029 | 0.029 | 0.029 | 0 |
Precision
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 |
Rate of recovery
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 | 87 |
| EDTA plasma (n=8) | 86-99 | 92 |
| Cell culture media (n=8) | 89-103 | 96 |
Linear
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 (%) | 86-101 | 88-97 | 89-97 |
| Average (%) | 91 | 96 | 92 | |
| 1:4 | Range (%) | 86-101 | 88-97 | 88-104 |
| Average (%) | 94 | 97 | 101 | |
| 1:8 | Range (%) | 87-96 | 91-103 | 87-96 |
| Average (%) | 92 | 96 | 93 | |
| 1:16 | Range (%) | 92-105 | 86-92 | 85-93 |
| Average (%) | 99 | 87 | 90 |
Assay Procedures
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