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Hamster TNF-a(Tumor Necrosis Factor-alpha) ELISA Kit
ELK0928
Size： Price：
48 T $265.00
96 T $376.00
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Overview

Product name: Hamster TNF-a(Tumor Necrosis Factor-alpha) ELISA Kit
Reactivity: Hamster
Alternative Names: DIF; TNF-A; TNFSF2; Cachectin; Tumor Necrosis Factor Ligand Superfamily Member 2
Assay Type: Sandwich
Sensitivity: 1.09 pg/mL
Standard: 200 pg/mL
Detection Range: 3.13-200 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;
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 Hamster TNF-a. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Hamster TNF-a. 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 Hamster TNF-a, 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 Hamster TNF-a in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
200.00 2.135 2.047
100.00 1.607 1.519
50.00 1.115 1.027
25.00 0.828 0.740
12.50 0.617 0.529
6.25 0.377 0.289
3.13 0.236 0.148
0.00 0.088 0.000

Precision

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.

Recovery

Matrices listed below were spiked with certain level of recombinant TNF-a and the recovery rates were calculated by comparing the measured value to the expected amount of TNF-a in samples.
Matrix Recovery range Average
serum(n=5) 78-95% 86%
EDTA plasma(n=5) 84-98% 91%
Heparin plasma(n=5) 85-97% 91%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of TNF-a and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Matrix 1:2 1:4 1:8 1:16
serum(n=5) 83-96% 79-93% 88-97% 89-102%
EDTA plasma(n=5) 83-97% 95-104% 88-104% 93-103%
Heparin plasma(n=5) 86-97% 83-96% 91-103% 89-96%