Rat V-Rel Reticuloendotheliosis Viral Oncogene Homolog A ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, listed under the Assays and Kits product category, for the in vitro quantitative determination of rat V-Rel Reticuloendotheliosis Viral Oncogene Homolog A (NF-kB p65) in tissue homogenates, cell lysates or other biological fluids. The kit shows reactivity with rat samples.

The assay uses a pre-coated 96-well microplate arranged as 12 x 8 well strips, with an antibody specific for NF-kB p65 immobilised on the plate. Standards and test samples are added to the wells and the target is bound by the immobilised antibody. Following incubation and washing, a biotinylated anti-NF-kB p65 detection antibody is added, then an HRP-Avidin conjugate. TMB substrate produces a blue-coloured product that changes to yellow upon addition of acidic stop solution; absorbance is read at 450 nm in a microplate reader and the NF-kB p65 concentration is calculated by reference to the standard curve. The reported assay range is 0.156-10 ng/ml with a sensitivity of <0.049 ng/ml and a total assay time of 4h 30m.

Recovery values in spiked samples (n=5): Serum 80-102% (average 91%), EDTA Plasma 81-100% (average 90%), Heparin Plasma 80-89% (average 84%). Linearity at dilutions of 1:2, 1:4, 1:8 and 1:16 (n=5): Serum 87-91%, 87-107%, 74-101%, 92-97%; EDTA Plasma 90-105%, 84-101%, 90-101%, 79-108%; Heparin Plasma 84-95%, 92-105%, 82-105%, 89-91%. Kit contents include pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard (2 vials), Detection Solution A (120 µl), Detection Solution B (120 µl), Wash Buffer 30X (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml) and Plate Sealers (5 adhesive strips). Components should be stored at +4°C or -20°C as instructed and the kit must not be used past its expiration date.