Mouse ALT ELISA Kit

A sandwich double-antibody enzyme-linked immunosorbent assay (ELISA) from Assay Genie, designed for the quantitative measurement of Mouse ALT — also known as alanine aminotransferase, alanine transaminase (ALAT), SGPT, or serum glutamic-pyruvic transaminase — in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The microplate is pre-coated with a capture antibody specific to the target analyte. Standards and appropriately diluted samples are incubated in the wells, followed by a biotinylated detection antibody specific to a different epitope of the protein, HRP-conjugated Streptavidin, and TMB substrate to generate a colorimetric reaction. The reaction is stopped with an acidic solution and absorbance is read at 450 nm using a microplate reader, with signal intensity proportional to analyte concentration as determined from a standard curve.

Linearity across dilution factors was 89–103% for serum (n = 5), 85–100% for EDTA plasma (n = 5), and 81–97% for heparin plasma (n = 5). Recovery ranges spanned 86–105% across the same matrices, with average recoveries of 94% for serum, 94% for EDTA plasma, and 97% for heparin plasma. The protocol calls for equilibrating reagents and TMB substrate to room temperature, then primary incubation at 37°C for 90 minutes, biotinylated detection antibody binding at 37°C for 60 minutes, HRP-Streptavidin binding at 37°C for 30 minutes, and TMB colour development in the dark for 10–20 minutes before reading absorbance at 450 nm. Reagents are stored at 2-8°C, with a stated shelf life of 12 months.