Rat Kidney Injury Molecule 1 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com for the in vitro quantitative determination of rat Kidney Injury Molecule 1 (TIM 1) in tissue homogenates, urine, cell culture supernates and other biological fluids. The assay uses a pre-coated 96-well microplate (12 x 8 well strips) bearing an antibody specific for TIM 1. Standards and test samples are added to the wells; after incubation and washing, a Biotinylated Anti-TIM 1 Antibody and then an HRP-Avidin conjugate are added. A TMB substrate visualises the HRP enzymatic reaction as a blue product that turns yellow on addition of an acidic stop solution. Absorbance is read at 450 nm in a microplate reader and TIM 1 concentration is calculated against the standard curve. Total assay time is 4 hours 30 minutes.

Sensitivity is reported as <22.6 pg/ml with an assay range of 62.5-4000 pg/ml, read on a colorimetric microplate reader. Components supplied include a pre-coated microplate (12 x 8 well strips, -20°C), lyophilised standard (2 vials, -20°C), Detection Solution A (120 μl, -20°C), Detection Solution B (120 μl, -20°C), 30X Wash Buffer (20 ml, +4°C), Sample Dilution Buffer (45 ml, -20°C), TMB Substrate (9 ml, +4°C), Stop Solution (6 ml, +4°C) and 5 adhesive plate sealers. Components should be stored at +4°C or -20°C as instructed and the kit should not be used past its expiration date.

Reported recovery values are: serum (n=5) 88-100%, average 94%; EDTA plasma (n=5) 90-101%, average 95%; heparin plasma (n=5) 92-101%, average 96%. Linearity across dilutions of 1:2, 1:4, 1:8 and 1:16 was demonstrated in serum (n=5) at 81-99%, 90-102%, 77-104% and 88-95%; in EDTA plasma (n=5) at 94-104%, 80-107%, 89-93% and 86-108%; and in heparin plasma (n=5) at 77-100%, 92-99%, 88-105% and 85-99%.