Rat Ghrelin ELISA Kit

Antibodies.com supplies a Rat Ghrelin ELISA Kit offered in two configurations: a competitive ELISA and a sandwich ELISA. Both are intended for the in vitro quantitative determination of rat Ghrelin in serum, plasma, tissue homogenates and related biological fluids (the competitive format additionally lists cell lysates and cell culture supernates), and both use a pre-coated 96-well microplate supplied as 12 x 8 well strips, read colorimetrically at 450 nm. The competitive configuration covers a product range of 0.156-10 ng/ml with a sensitivity of <0.042 ng/ml and a total assay time of 2 h 30 min; the sandwich configuration covers a product range of 7.813-500 pg/mL with a sensitivity of <4.688 pg/mL and a total assay time of 4 hours.

In the competitive assay, Ghrelin in samples or standards competes with Ghrelin pre-coated on the plate for binding to a biotinylated anti-Ghrelin antibody, and detection proceeds via an HRP-avidin conjugate and TMB substrate, producing a yellow colour inversely proportional to Ghrelin concentration. In the sandwich assay, Ghrelin is captured on an immobilised anti-Ghrelin antibody and detected with a biotinylated anti-Ghrelin antibody plus HRP-Streptavidin, producing a yellow colour directly proportional to Ghrelin concentration. The sandwich configuration is described as specific for Ghrelin with no reported cross-reactivity with other analogs or variants.

Both formats have been validated in serum, EDTA plasma and heparin plasma. Reported recovery values are, for the competitive format: serum average 91% (range 80%-102%, n=5), EDTA plasma 90% (81%-100%, n=5) and heparin plasma 84% (80%-89%, n=5); and for the sandwich format: serum 97% (92%-101%, n=10), EDTA plasma 95% (87%-104%, n=10) and heparin plasma 98% (93%-105%, n=10). For the sandwich format, serum and plasma samples should be diluted at least 1:2 with Sample Dilution Buffer prior to testing, with validated dilutions of 1:5-1:20 for healthy serum and plasma; users are advised to avoid freeze/thaw cycles. Kit components include a pre-coated microplate, lyophilised standard, detection reagents, wash buffer, sample dilution buffer, TMB substrate, stop solution and plate sealers; components should be stored at +4°C or -20°C as instructed, and the kit must not be used past its expiration date.