Mouse Wingless Type MMTV Integration Site Family, Member 5A ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) supplied by Antibodies.com is intended for the in vitro quantitative determination of mouse Wnt5a (Wingless Type MMTV Integration Site Family, Member 5A) in tissue homogenates, cell lysates, cell culture supernates and other biological fluids. The assay uses a pre-coated 96-well microtiter plate arranged as 12 x 8 well strips, on which an antibody specific for Wnt5a has been immobilised.

Standards and test samples are added to the wells so that Wnt5a is captured by the immobilised antibody. After incubation and washing, a Biotinylated Anti-Wnt5a Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate solution is then used to visualise the HRP enzymatic reaction; the resulting blue product turns yellow upon addition of acidic stop solution, and the concentration of Wnt5a is calculated from the absorbance at 450 nm against the standard curve.

Quantitative performance is supported by a product range of 31.2–2000 pg/ml and a sensitivity of <12.4 pg/ml, with a total assay time of 4 h 30 min. Average recovery across tested samples was 91% for serum (range 83–100%, n=5), 94% for EDTA plasma (range 90–99%, n=5) and 96% for heparin plasma (range 91–101%, n=5), and dilution linearity has been demonstrated from 1:2 through 1:16 for serum, EDTA plasma and heparin plasma. Kit components are stored at +4°C or −20°C as instructed, and the kit must not be used past its expiration date.