Rat Microfibrillar Associated Protein 4 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of rat Microfibrillar Associated Protein 4 (MFAP4) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates, or other biological fluids. The assay employs the sandwich enzyme immunoassay technique, in which an antibody specific for MFAP4 is pre-coated onto a 96-well microtiter plate. Standards and test samples are added to the wells, allowing MFAP4 to bind the immobilised antibody. After incubation and washing, a Biotinylated Anti-MFAP4 Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate solution then visualises the HRP enzymatic reaction, producing a blue colour that changes to yellow upon addition of acidic stop solution. The intensity of the yellow colour is proportional to the amount of MFAP4 captured, and the concentration is calculated by reading the O.D. absorbance at 450 nm in a microplate reader and referring to the standard curve.

Key specifications include a sensitivity of 0.115 ng/ml, a measuring range of 0.312–20 ng/ml, and detection via a colorimetric microplate reader on a pre-coated 96-well microplate platform arranged as 12 x 8 well strips. The total assay time is 4h 30m. Storage instructions indicate that kit components should be stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.

Recovery performance was evaluated across sample types: serum (n=5) showed an average of 91% (range 80–102%); EDTA plasma (n=5) showed an average of 90% (range 81–100%); and heparin plasma (n=5) showed an average of 84% (range 80–89%). Dilution linearity was also reported. At 1:2, 1:4, 1:8, and 1:16 dilutions, serum (n=5) ranged 87–91%, 87–107%, 74–101%, and 92–97%; EDTA plasma (n=5) ranged 90–105%, 84–101%, 90–101%, and 79–108%; and heparin plasma (n=5) ranged 84–95%, 92–105%, 82–105%, and 89–91%. Kit components include a Pre-Coated 96 Well Microplate, Lyophilized Standard, Detection Solution A, Detection Solution B, Wash Buffer (30X), Sample Dilution Buffer, TMB Substrate, Stop Solution, and Plate Sealers.