Mouse Cyclin Dependent Kinase Inhibitor 2A ELISA Kit

A sandwich ELISA from Antibodies.com for the in vitro quantitative determination of mouse CDKN2A in tissue homogenates or other biological fluids. The kit uses the sandwich enzyme immunoassay technique: an antibody specific for CDKN2A is pre-coated onto a 96-well microplate, and CDKN2A in standards or test samples is captured by the immobilised antibody. After washing, a Biotinylated Anti-CDKN2A Antibody binds the captured analyte, followed by an HRP-Avidin conjugate. TMB substrate is then added; HRP catalyses conversion to a blue product that turns yellow upon addition of acidic stop solution. The intensity of the yellow colour is proportional to the amount of CDKN2A captured, and concentration is calculated by reading O.D. absorbance at 450 nm against the standard curve.

Reported sensitivity is <0.108 ng/ml with an assay range of 0.312–20 ng/ml, and the total assay time is 4 h 30 min. Detection is colorimetric on a pre-coated microplate of 12 × 8 well strips. Spike-and-recovery data (n=5) give averages of 91% for serum (range 83–100%), 94% for EDTA plasma (range 90–98%) and 94% for heparin plasma (range 90–99%). Dilution linearity at 1:2, 1:4, 1:8 and 1:16 is reported for serum, EDTA plasma and heparin plasma samples. The protocol is described as easy to follow.

Kit components include a pre-coated 96-well microplate, lyophilised standard, Detection Solution A, Detection Solution B, 30× Wash Buffer, Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers. Components are stored at +4°C or –20°C as instructed, and the kit must not be used past its expiration date.