Porcine Connexin 43 ELISA Kit

From Antibodies.com, this Sandwich ELISA kit enables in vitro quantitative determination of porcine Connexin 43/GJA1 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. The assay uses the sandwich enzyme immunoassay technique, with an antibody specific for Connexin 43/GJA1 pre-coated onto a 96-well microtiter plate.

Following incubation of standards and test samples, wells are washed and incubated with a Biotinylated Anti-Connexin 43/GJA1 Antibody, then with an HRP-Avidin conjugate. TMB substrate solution visualises the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of an acidic stop solution. Absorbance is read at 450 nm in a colorimetric microplate reader, and the concentration of Connexin 43/GJA1 is calculated against the standard curve. The reported working range is 0.625–40 ng/ml, with a sensitivity of <0.16 ng/ml, and a total assay time of 4h 30m.

Reported spike-by-sword recovery values are serum (n=5) 80–102% (average 91%), EDTA plasma (n=5) 81–100% (average 90%), and heparin plasma (n=5) 80–89% (average 84%). Dilution linearity at 1:2, 1:4, 1:8 and 1:16 respectively yields: serum (n=5) 87–91%, 87–107%, 74–101%, 92–97%; EDTA plasma (n=5) 90–105%, 84–101%, 90–101%, 79–108%; heparin plasma (n=5) 84–95%, 92–105%, 82–105%, 89–91%. Kit contents include a pre-coated 96-well microplate (12 × 8 well strips), lyophilised standard, Detection Solution A, Detection Solution B, Wash Buffer (30X), Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers. Kit components should be stored at +4°C or -20°C as instructed and the kit must not be used past its expiration date.