Mouse Tumor Necrosis Factor Receptor Superfamily, Member 5 ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of mouse CD40 (Tumor Necrosis Factor Receptor Superfamily, Member 5) in serum, plasma or other biological fluids. The kit employs the sandwich enzyme immunoassay technique, with an antibody specific for CD40 pre-coated onto a 96-well microtiter plate. Standards and test samples are added to the wells, where CD40 present in each sample is bound by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-CD40 Antibody is added, followed by HRP-Avidin conjugate. TMB substrate solution is then used to visualise the HRP enzymatic reaction by catalysis, producing a blue-coloured product that changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of CD40 captured in each well, and the concentration of CD40 is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

The assay has a sensitivity of <5.4 pg/ml and an assay range of 15.6–1000 pg/ml. Recovery has been validated in serum (range 80%–102%, average 91%, n=5), EDTA plasma (range 81%–101%, average 91%, n=5) and heparin plasma (range 80%–89%, average 84%, n=5). Dilution linearity was also assessed at 1:2, 1:4, 1:8 and 1:16 dilutions across serum, EDTA plasma and heparin plasma samples. The total assay time is approximately 4 hours 30 minutes, and detection is colourimetric on a pre-coated microplate platform. Kit components should be stored at +4°C or –20°C as instructed, and the kit should not be used past its expiration date.