Human Kidney Injury Molecule 1 ELISA Kit

A sandwich enzyme-linked immunosorbent assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of human Kidney Injury Molecule 1 (TIM 1) in tissue homogenates, urine, cell culture supernates, or other biological fluids. The test uses a pre-coated microplate with a capture antibody specific for TIM 1 immobilised on the plate. Standards and test samples are added and any TIM 1 present is bound by the capture antibody. After washing, a biotinylated anti-TIM 1 antibody is added, followed by an HRP-avidin conjugate. TMB substrate produces a blue colour that changes to yellow upon addition of the acidic stop solution. The intensity of yellow is proportional to the TIM 1 concentration in the well, which is calculated from the O.D. absorbance at 450 nm in a colorimetric microplate reader against the standard curve.

The standard curve covers 31.2–2000 pg/ml with a sensitivity of <11.6 pg/ml. Reported recovery in human samples (n=5) ranged from 88% to 100% in serum, 90% to 101% in EDTA plasma, and 92% to 101% in Heparin plasma. Linearity was demonstrated across 1:2 to 1:16 dilutions with recoveries of 77–104% in serum, 80–108% in EDTA plasma, and 77–105% in Heparin plasma (all n=5). Total assay time is approximately 4 hours 30 minutes.

Components are stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date. Reagents include a pre-coated microplate, lyophilised standard, detection reagents, wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers. The protocol is straightforward to follow.