Human Thyroid Hormone Receptor Alpha ELISA Kit

This Antibodies.com kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Thyroid Hormone Receptor Alpha in tissue homogenates or other biological fluids. The assay uses a pre-coated microtiter plate with an antibody specific for Thyroid Hormone Receptor Alpha. Standards and samples are added to the wells, where the target analyte is captured by the immobilized antibody. Following incubation and washing, a Biotinylated Anti-Thyroid Hormone Receptor Alpha Antibody is added, followed by HRP-Avidin conjugate and further incubation. TMB substrate solution is then used to visualize the HRP enzymatic reaction, producing a blue product that changes to yellow upon addition of acidic stop solution. Absorbance is read at 450 nm in a microplate reader, and concentration is calculated by reference to a standard curve.

Validated for human reactivity, the assay has a quantitative range of 62.5-4000 pg/ml and a sensitivity of 22.5 pg/ml, with a total assay time of 4 hours 30 minutes. Sample types include tissue homogenates and other biological fluids. Spike-and-recovery testing (n=5) yielded average recoveries of 91% in serum (range 80-102%), 90% in EDTA plasma (range 81-100%), and 84% in heparin plasma (range 80-89%). Linearity at 1:2, 1:4, 1:8, and 1:16 dilutions showed serum recoveries of 87-91%, 87-107%, 74-101%, and 92-97%; EDTA plasma of 90-105%, 84-101%, 90-101%, and 79-108%; and heparin plasma of 84-95%, 92-105%, 82-105%, and 89-91%.

Kit components are stored at +4°C or -20°C as instructed in the protocol manual, and the kit should not be used past its expiration date. Detection is colorimetric and performed on a microplate reader.