Human Occludin ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, designed for the in vitro quantitative determination of human Occludin in tissue homogenates, cell lysates or other biological fluids. The assay uses a 96-well microplate pre-coated with an antibody specific for Occludin. Standards and samples are added to the wells and bound by the immobilised antibody; following incubation and washing, a Biotinylated Anti-Occludin Antibody is added, followed by an HRP-Avidin conjugate. After further incubation and washing, TMB substrate is used to visualise the HRP enzymatic reaction, producing a blue product that turns yellow upon addition of stop solution. The density of yellow is proportional to the amount of Occludin captured, and concentration is calculated from the O.D. absorbance read at 450nm by reference to a standard curve.

The kit has a stated assay range of 0.156–10 ng/ml, a sensitivity of <0.058 ng/ml, and a total assay time of 4 hours 30 minutes. Recovery has been characterised in serum, EDTA plasma and heparin plasma samples at multiple dilution factors. The kit is suitable for use with cell lysates and tissue homogenates. Detection is colorimetric, and the kit is intended for use with a colorimetric microplate reader.

Components supplied include a pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard, detection solution A, detection solution B, 30X wash buffer, sample dilution buffer, TMB substrate, stop solution, and plate sealers, with storage conditions specified per component at +4°C or -20°C. Kit components should not be used past their expiration date.