Rat Tumor Necrosis Factor Beta ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay from Antibodies.com for the in vitro quantitative determination of rat Tumor Necrosis Factor Beta (also known as LTA) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates, and other biological fluids. The assay uses a pre-coated microplate on which standards and samples are added; LTA in the sample binds the immobilised capture antibody, followed by sequential incubation with a biotinylated anti-LTA detection antibody and an HRP-avidin conjugate. TMB substrate is then added to visualise the enzymatic reaction, producing a colour change proportional to bound analyte, and the LTA concentration is determined from the optical density read at 450 nm on a colorimetric microplate reader.

The standard curve spans 15.6–1000 pg/ml with a sensitivity of 5.7 pg/ml, and the full protocol runs in 4 hours 30 minutes. In spike-and-recovery testing, average recoveries were 91% in serum (range 80–102%, n=5), 90% in EDTA plasma (range 81–100%, n=5), and 84% in heparin plasma (range 80–89%, n=5), with dilution linearity demonstrated for each matrix at 1:2, 1:4, 1:8 and 1:16 sample dilutions. Components supplied include a pre-coated microplate, lyophilised standard, detection solutions A and B, wash buffer, sample dilution buffer, TMB substrate, stop solution and adhesive plate sealers, which should be stored at +4°C or −20°C as indicated on the individual components; the kit should not be used past its expiration date.