Mouse Inter Alpha-Globulin Inhibitor H4 ELISA Kit

This sandwich enzyme-linked immunosorbent assay (sELISA), supplied by Antibodies.com, is designed for the in vitro quantitative determination of mouse Inter Alpha-Globulin Inhibitor H4 (ITIH4) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. The assay employs a pre-coated 96-well microplate arranged as 12 x 8 well strips, onto which an antibody specific for ITIH4 has been immobilised. Standards and test samples are added to the wells, and ITIH4 present in each sample binds to the immobilised antibody. After washing, a Biotinylated Anti-ITIH4 Antibody binds the captured ITIH4, followed by HRP-Avidin conjugate. TMB substrate solution is then added; catalysis of the HRP reaction produces a blue-coloured product that changes to yellow upon addition of acidic stop solution. The intensity of yellow is proportional to the amount of ITIH4 captured, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a microplate reader against the standard curve.

Key specifications: reactivity — Mouse; protein target — ITIH4; assay type — sandwich (quantitative); detection type — colorimetric; assay time — 4h 30m; product range — 78.1–5000 ng/ml; sensitivity — <33 ng/ml; instrument — colorimetric microplate reader; storage — store kit components at +4 °C or −20 °C as instructed; do not use past the expiration date.

Spike-and-recovery validation (n=5) gave averages of 91% for serum (range 80–102%), 90% for EDTA plasma (range 81–100%) and 84% for heparin plasma (range 80–89%). Linearity across dilution series 1:2, 1:4, 1:8 and 1:16 (n=5) showed serum recovery of 87–91%, 87–107%, 74–101% and 92–97%; EDTA plasma at 90–105%, 84–101%, 90–101% and 79–108%; and heparin plasma at 84–95%, 92–105%, 82–105% and 89–91%.