Human High Mobility Group Protein 1 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) supplied by Antibodies.com for the in vitro quantitative determination of human HMGB1 in serum, plasma, or other biological fluids. The assay uses a pre-coated 96-well microplate arranged as 12 x 8 well strips, with an antibody specific for HMGB1 immobilised on the plate. Standards and samples are added to the wells and HMGB1 is captured by the immobilised antibody; after washing, a biotinylated anti-HMGB1 detection antibody binds the captured analyte, followed by an HRP-Avidin conjugate and TMB substrate. The enzymatic reaction produces a colour change whose intensity is read as absorbance at 450 nm against a standard curve.

The reported assay range is 62.5–4000 pg/ml and the sensitivity is <23.7 pg/ml. Detection is colorimetric and the assay is intended to be read on a microplate reader. The total assay time is 4h 30m.

Spike-and-recovery has been characterised for serum (n=5; range 80%–102%; average 91%), EDTA plasma (n=5; range 81%–100%; average 90%) and heparin plasma (n=5; range 80%–89%; average 84%), with dilution linearity also reported across 1:2, 1:4, 1:8 and 1:16 dilutions for the same sample types. Kit components are shipped at +4°C and stored at +4°C or −20°C as instructed, and the kit should not be used past its expiration date.