Rat Cathepsin B ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of rat Cathepsin B in serum, plasma or other biological fluids.

The kit employs the sandwich enzyme immunoassay technique. An antibody specific for Cathepsin B is pre-coated onto a microplate. Standards and test samples are added to the wells and Cathepsin B present in each sample is captured by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-Cathepsin B Antibody binds the captured Cathepsin B in each well. An HRP-Avidin conjugate is then added and the plate is incubated. After further washing, TMB substrate solution is used to visualise the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of Cathepsin B captured in each well. The concentration of Cathepsin B is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

Reported assay range is 0.156–10 ng/ml with sensitivity of 0.062 ng/ml. Average recovery values were 91% in serum (range 80–102%, n=5), 90% in EDTA plasma (range 81–100%, n=5) and 84% in heparin plasma (range 80–89%, n=5). Dilution linearity data are provided for serum, EDTA plasma and heparin plasma at dilutions of 1:2, 1:4, 1:8 and 1:16. Total assay time is 4h 30m. Detection is colorimetric, read on a microplate reader, and kit components are stored at +4°C or –20°C as instructed.