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Product name: | Rat TNFa(Tumor Necrosis Factor Alpha) ELISA Kit |
Reactivity: | Rat |
Alternative Names: | TNF Alpha; DIF; TNF-A; TNFSF2; Cachectin; Tumor Necrosis Factor Ligand Superfamily Member 2 |
Assay Type: | Sandwich |
Sensitivity: | 6.1 pg/mL |
Standard: | 1000 pg/mL |
Detection Range: | 15.63-1000 pg/mL |
Sample Type: | serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
Assay Length: | 3.5h |
Research Area: | Cytokine;Tumor immunity;Infection immunity; |
Uniprot ID: | P16599 |
Test principle: | 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 Rat TNFa. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat TNFa. 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 Rat TNFa, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rat TNFa in the samples is then determined by comparing the OD of the samples to the standard curve. |
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Concentration (pg/mL) | OD | Corrected OD |
---|---|---|
1000.00 | 2.345 | 2.256 |
500.00 | 1.769 | 1.680 |
250.00 | 1.225 | 1.136 |
125.00 | 0.917 | 0.828 |
62.50 | 0.521 | 0.432 |
31.25 | 0.366 | 0.277 |
15.63 | 0.207 | 0.118 |
0.00 | 0.089 | 0.000 |
Intra-assay Precision (Precision within an assay):CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays):CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Matrix | Recovery range | Average |
---|---|---|
serum(n=5) | 80-90% | 85% |
EDTA plasma(n=5) | 84-106% | 100% |
Heparin plasma(n=5) | 87-93% | 90% |
Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
---|---|---|---|---|
serum(n=5) | 89-97% | 85-94% | 87-96% | 85-97% |
EDTA plasma(n=5) | 85-94% | 87-96% | 92-101% | 82-98% |
Heparin plasma(n=5) | 85-99% | 98-106% | 86-99% | 78-95% |
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