Interferon gamma (IFN-gamma) BioAssay™ ELISA Kit (Hamster)

The Interferon gamma (IFN-gamma) BioAssay™ ELISA Kit is a quantitative sandwich assay for the detection of IFN-gamma in hamster serum, plasma, tissue homogenates and other biological fluids.

Detection Range:
15.625-1000pg/ml

Sensitivity:
9.375pg/ml

Precision:
Intra-Assay CV: <8%
Inter-Assay CV: <10%

Assay Principle:
The microtiter plate provided in this kit has been pre-coated with an antibody specific to IFN-gamma. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to IFN-gamma. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain IFN-gamma, biotin-conjugated antibody and enzyme-conjugated streptavidin complex will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of IFN-gamma in the sample is then determined by comparing the O.D. of the sample to the standard curve.

Kit Components:
369991A: Microtiter Strips, 1x48 wells or 1x96 wells
369991B: Standard, 1x1vial or 2x1vial
369991C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml
369991D: Antibody (Biotin) (Concentrated), 1x60ul or 1x120ul
369991E: Antibody Dilution Buffer, 1x5ml or 1x10ml
369991F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul
369991G: SABC Dilution Buffer, 1x5ml or 1x10ml
369991H: TMB Substrate, 1x5ml or 1x10ml
369991J: Stop Solution, 1x5ml or 1x10ml
369991K: Wash Buffer, 25X, 1x15ml or 1x30ml

Storage and Stability:
Store unopened *369991A at 4ºC; store at -20ºC once opened. Store unopened *369991B at 4°C; once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.

Assay Summary:
1. Wash plate 2 times before adding standards, samples and control (zero) to wells.
2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C. Aspirate and wash 2 times.
3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 minutes at 37°C
4. Aspirate and wash 3 times.
5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C
6. Aspirate and wash 5 times.
7. Add 90ul TMB substrate. Incubate 10-20 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
9. Calculate results.