Human TSH ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human TSH in serum, plasma, and cell culture supernatant.
Assay Type
Sandwich (quantitative)
Principle of Assay
Human TSH ELISA Kit (A334835) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human TSH in serum, plasma, and cell culture supernatant.. An antibody specific for TSH has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the TSH present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-TSH Antibody, which binds the captured TSH present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Streptavidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of TSH captured in each well. The concentration of TSH can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
Components
This ELISA Kit contains 10X 600 µL Capture Antibodies, 10X 600 µL Detector Antibodies, 2 Lyophilized Recombinant Protein Vials, 30 mL Assay Diluent #12Y, 10X 20 mL Wash Buffer, 12 mL TMB Solution, 12 mL Stop Solution, Pre-Coated 96-Well H-Microplate (12 × 8 well strips), and 2 Plate Seals.
The standard curve for human thyroid-stimulating hormone (TSH) was generated by plotting optical density (O.D. 450 nm) versus TSH concentration (pg/mL) in Assay Diluent #12Y on a log-log scale. The curve demonstrates a strong linear relationship across the tested range, indicating reliable assay sensitivity and quantification accuracy for TSH detection.
The standard curve for human thyroid-stimulating hormone (TSH) was established by plotting optical density (O.D. 450 nm) against TSH concentration (µIU/mL) in Assay Diluent #12Y on a log-log scale. The curve shows a strong linear correlation across the tested range, demonstrating the assay’s accuracy and sensitivity for quantifying TSH levels.
Human serum samples were analyzed for thyroid-stimulating hormone (TSH) concentrations using an ELISA assay at different dilution factors (undiluted, 2×, 4×, and 8×). The results show consistent detection of TSH across dilutions, with minimal variation, demonstrating the assay’s robustness and dilution linearity for serum samples. Data are expressed as mean ± SD.
Human plasma samples were tested for thyroid-stimulating hormone (TSH) concentrations using an ELISA at different dilution factors (undiluted, 2×, 4×, 8×, and 16×). The results demonstrate consistent TSH detection across dilutions, indicating good assay performance and dilutional linearity in plasma matrices. Data are presented as mean ± SD.
Percentage recovery of human TSH was assessed at various dilution levels (13%, 6%, 3%, 2%, and 1%) for plasma (brown line) and cell culture media (blue line). Both sample types showed acceptable recovery across dilutions, with plasma demonstrating slightly higher recovery consistency compared to cell culture media.
Percentage recovery of human TSH was evaluated at different dilution levels (10.0%, 5.0%, 2.5%, 1.3%, and 0.6%) in serum samples. Recovery values remained consistent across dilutions, indicating reliable assay performance and minimal matrix interference.
