Rat Lecithin Cholesterol Acyltransferase ELISA Kit

The Antibodies.com Rat LCAT ELISA Kit is a sandwich enzyme-linked immunosorbent assay (sELISA) designed for the in vitro quantitative determination of rat Lecithin Cholesterol Acyltransferase (LCAT) in serum, plasma or other biological fluids. The assay employs a 96-well microplate pre-coated with an antibody specific for LCAT. Standards and test samples are added to the wells, where LCAT present in each sample is captured by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-LCAT Antibody is added and binds the captured LCAT. An HRP-Avidin conjugate is then added and incubated, followed by TMB substrate solution which produces a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of LCAT captured in each well, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a microplate reader against a standard curve.

The kit has an assay range of 0.156-10 ng/ml and a sensitivity of <0.052 ng/ml. The total assay time is 4h 30m and the assay is read on a colourmetric microplate reader. Spike-and-recovery testing yielded average recoveries of 91% for serum (range 80% - 102%, n=5), 90% for EDTA plasma (range 81% - 100%, n=5) and 84% for heparin plasma (range 80% - 89%, n=5). Dilution linearity was tested at 1:2, 1:4, 1:8 and 1:16 dilutions (all n=5): serum ranged from 87-91% to 92-97%, EDTA plasma ranged from 90-105% to 79-108%, and heparin plasma ranged from 84-95% to 89-91%. Kit components should be stored at +4°C or -20°C as instructed and not used past the expiration date.