Human Connexin 40 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) kit from Antibodies.com designed for the in vitro quantitative determination of human Cx40/GJA5 in tissue homogenates or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, with an antibody specific for Cx40/GJA5 pre-coated onto a 96-well microplate. Standards and test samples are added to the wells, where Cx40/GJA5 is captured by the immobilised antibody, followed by incubation with a Biotinylated Anti-Cx40/GJA5 Antibody and an HRP-Avidin conjugate. A TMB substrate produces a blue-coloured product that changes to yellow after addition of acidic stop solution; the density of yellow is proportional to the amount of Cx40/GJA5 captured and the concentration is calculated from O.D. absorbance at 450 nm in a microplate reader by reference to a standard curve.

The kit is supplied as a pre-coated microplate of 12 x 8 well strips and is read colourimetrically, with a total assay time of 4h 30m. The manufacturer reports a sensitivity of <0.103 ng/ml and an assay range of 0.312-20 ng/ml. Mean recovery values (n=5) are stated as 91% for serum (range 83–100%), 94% for EDTA plasma (range 91–98%) and 94% for heparin plasma (range 90–99%). For sample dilutions of 1:2, 1:4, 1:8 and 1:16 (n=5), reported ranges are 81–110%, 82–98%, 90–100% and 80–108% for serum; 91–97%, 90–107%, 70–108% and 89–93% for EDTA plasma; and 86–104%, 79–103%, 95–99% and 86–110% for heparin plasma. Kit components should be stored at +4°C or -20°C as instructed, and the kit must not be used past the expiration date.