The Interleukin-12 subunit alpha (IL12A) BioAssay™ ELISA Kit (Human) is a sandwich ELISA for the quantitative determination of human interleukin-12 subunit alpha (IL12A) concentrations in serum, plasma, cell culture supernates.
Sensitivity:3.9pg/ml
Detection Range:15.6-1000pg/ml.
Specificity:This assay has high sensitivity and excellent specificity for detection of human IL12A. No significant cross-reactivity or interference between human IL12A and analogues was observed.
Precision:Intra-assay: CV%<8%Inter-assay: CV%<10%
Test Principle:An antibody specific for IL12A has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL12A present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for IL12A 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 IL12A bound in the initial step. The color development is stopped and the intensity 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.
