Human NT proBNP ELISA Kit

From Assay Genie, this sandwich double-antibody enzyme-linked immunosorbent assay (ELISA) enables quantitative measurement of human N-terminal pro-Brain Natriuretic Peptide (NT-proBNP). The microplate is pre-coated with a capture antibody specific to the target protein; standards and appropriately diluted samples are added and incubated, allowing the analyte to bind. A biotinylated detection antibody directed against a separate epitope then forms a sandwich complex, followed by HRP-conjugated Streptavidin and TMB substrate to produce a colorimetric reaction. The reaction is stopped with an acidic solution and absorbance is read at 450 nm.

Validated for human reactivity, the kit accepts serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples, and should be stored at 2-8°C for 12 months. The reported sensitivity is 23.438 pg/mL and the assay range is 39.063-2500 pg/mL. Linearity across 1:2, 1:4 and 1:8 dilutions was 87-105%, 87-97% and 85-100% for serum (n=5); 93-100%, 83-98% and 85-98% for EDTA plasma (n=5); and 88-98%, 82-94% and 82-96% for heparin plasma (n=5). Average recovery was 96% for serum (range 88-101%), 98% for EDTA plasma (range 92-104%) and 95% for heparin plasma (range 87-99%).

The procedure involves equilibrating reagents to room temperature, loading standards, samples and blanks onto the pre-coated plate and incubating at 37°C for 90 minutes. A biotinylated detection antibody is then added for a 60-minute incubation, followed by HRP-Streptavidin (SABC) for 30 minutes, TMB substrate development in the dark for 10-20 minutes, addition of stop solution, and immediate absorbance measurement at 450 nm.