Rat High Density Lipoprotein ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (ELISA) from Antibodies.com enables the in vitro quantitative determination of rat high density lipoprotein (HDL) in serum, plasma or other biological fluids. The procedure uses a 96-well microtiter plate pre-coated with an antibody specific for HDL. Standards and samples are added to the wells where HDL is captured by the immobilized antibody. After washing, a Biotinylated Anti-HDL Antibody is added, followed by HRP-Avidin conjugate. TMB substrate solution produces a blue-coloured product that turns yellow after addition of acidic stop solution, with absorbance read at 450 nm to calculate HDL concentration against a standard curve.

The assay provides a sensitivity of <1.39 ng/ml across an assay range of 3.13-200 ng/ml and is read on a colorimetric microplate reader. Total assay time is 4h 30m. Validation in serum (n=5) yielded recoveries of 80-102% with an average of 91%, EDTA plasma (n=5) recoveries of 81-100% averaging 90%, and heparin plasma (n=5) recoveries of 80-89% averaging 84%, with recommended sample dilutions described in the protocol. Kit components include a pre-coated 96-well microplate arranged as 12 x 8 well strips, lyophilized standard, detection solutions, wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers, stored at +4°C or -20°C as instructed.