Rat Substance P ELISA Kit

A competitive Enzyme-Linked Immunosorbent Assay (cELISA) from Antibodies.com, designed for the in vitro quantitative determination of rat Substance P in serum, plasma and other biological fluids. The assay employs the competitive enzyme immunoassay technique, using a microtiter plate pre-coated with Substance P antigen. During incubation, Substance P present in samples or standards competes with the fixed amount of immobilized antigen for binding sites on a Biotinylated Anti-Substance P Antibody. Unbound antibody is removed by washing, and an HRP-Avidin conjugate is added to the wells. TMB substrate solution is then used to visualize the HRP enzymatic reaction, producing a blue colour that changes to yellow upon addition of acidic stop solution. The density of yellow is inversely proportional to the amount of Substance P in each sample or standard.

The kit has a reported sensitivity of 4.63 pg/ml and an assay range of 15.6-1000 pg/ml. Detection is colorimetric, with the concentration of Substance P calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve. The total assay time is 2h 30m, and kit components should be stored at +4°C or -20°C as instructed. The kit should not be used past its expiration date.

Recovery has been characterised in serum (range 80%-102%, average 91%), EDTA plasma (range 81%-99%, average 90%) and heparin plasma (range 80%-89%, average 84%). Dilution linearity is also reported across dilutions of 1:2, 1:4, 1:8 and 1:16 for serum, EDTA plasma and heparin plasma.