The Interferon γ (IFN-γ) BioAssay™ ELISA Kit (Human) is a quantitative sandwich ELISA for the quantitative determination of human interferon γ (IFN-γ) concentrations in serum, cell culture supernates, urine, tissuehomogenates and cerebrospinal fluid (CSF).
Sensitivity:1.56pg/ml
Detection Range:6.25-400pg/ml.
Specificity:This assay has high sensitivity and excellent specificity for detection of human IFN-γ. No significant cross-reactivity or interference between human IFN-γ and analogues was observed.
Precision:Intra-assay: CV%<8%Inter-assay: CV%<10%
Test Principle:An antibody specific for IFN-γ has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IFN-γ present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for IFN-γ is added to the wells. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IFN-γ bound in the initial step. The color development is stopped and theintensity of the color is measured.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Assay Summary:1. Prepare reagents, standards and samples as instructed
2. Add 100ul standard or sample to each well. Incubate for 1 hour at 37°C3. Aspirate and wash 3 times
4. Add 100ul MAU/ALB (Biotin), 1X to each well. Incubate for 1 hour at 37°C5. Aspirate and wash 3 times
6. Add 100ul Avidin (HRP), 1X to each well. Incubate for 1 hour at 37°C7. Aspirate and wash 5 times
8. Add 90ul TMB substrate to each well Incubate for 20 minutes at 37°C. Protect from light9. Add 50ul Stop Solution to each well. Read at 450nm within 5 minutes.
