Mouse HMGB1 ELISA Kit

Assay Genie offers a Sandwich ELISA, Double Antibody kit designed for the quantitative measurement of mouse HMGB1 (High Mobility Group Box 1) in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The colorimetric workflow uses a pre-coated capture microplate, a biotinylated detection antibody directed to a different epitope of the target, HRP-conjugated Streptavidin, and TMB substrate, with absorbance read at 450 nm. The kit is supplied for use within immunology research.

Reported performance characteristics include a sensitivity of 9.375 pg/mL and a standard curve range of 15.625-1000 pg/mL. Linearity across dilutions of 1:2, 1:4 and 1:8 is 92-100%, 88-105% and 89-105% for serum (n = 5); 86-99%, 82-90% and 87-96% for EDTA plasma (n = 5); and 84-100%, 87-99% and 80-104% for heparin plasma (n = 5). Recovery ranges are 95-105% (average 97%) for serum, 86-101% (average 95%) for EDTA plasma, and 92-103% (average 93%) for heparin plasma. The kit is stored at 2-8°C for 12 months. Listed synonyms for the analyte include Hmgb1, HMG-1, Amphoterin, High mobility group protein 1, HMG3, SBP-1, and Sulfoglucuronyl carbohydrate binding protein.

The assay procedure involves equilibrating reagents and the TMB substrate to room temperature, loading standards and samples onto the coated plate and incubating at 37°C for 90 minutes, adding biotin-labeled detection antibody for a 60-minute incubation at 37°C, adding HRP-Streptavidin for 30 minutes, developing with TMB substrate in the dark for 10-20 minutes, and reading absorbance at 450 nm immediately after adding the stop solution.