TSH (Mouse) ELISA

Enzyme immunoassay for the quantitative measurement of TSH in mouse serum and EDTA plasma. For research use only, not for use in diagnostic procedures.

Thyroid-stimulating hormone (also known as thyrotropin or TSH) is a glycoprotein produced by the anterior pituitary gland. Through its action on the thyroid gland, it plays a major role in maintaining normal circulating levels of the iodothyronines, T4 and T3. The production and secretion of TSH is controlled on the side by negative feedback from circulating T4 and T3, and on the other side by the hypothalamic thyrotropin-releasing hormone (TRH). The TSH molecule is composed of two non-identical subunits, α and β, that are bound together in a noncovalent manner. Within a species, the TSH α subunit is structurally identical to the α subunits of related glycoprotein hormones (LH, FSH). The β subunits of the related hormones are structurally hormone-specific and therefore determine their unique biological activities. Mouse models are widely accepted and indispensable for studying thyroid-stimulating hormone (TSH) regulation within the hypothalamic-pituitary-thyroid axis, owing to the high conservation of TSH signaling and feedback mechanisms between mice and humans. Genetic and disease-oriented mouse models have been essential for elucidating both fundamental TSH physiology and TSH-driven disorders (1–3).

The TSH mouse ELISA is a solid phase enzyme-linked immunosorbent assay (ELISA) in the microtiter plate format, designed for the quantitative measurement of TSH in mouse serum and EDTA plasma. The microtiter plate is coated with a monoclonal antibody directed against TSH β subunit. Calibrators and samples are pipetted in the coated wells and incubated with the enzyme conjugate (horseradish peroxidase-labeled TSH antibody). During 18-24 hours incubation at 2-8°C sandwich complexes consisting of the two antibodies and the mouse TSH are formed. Unbound components are removed by a washing step. Subsequently, the chromogenic substrate TMB (3,3',5,5'-tetramethyl-benzidine), is added to all wells. During a 30 minutes incubation, the substrate is converted to a colored end product (blue) by the bound enzyme. Enzyme reaction is stopped by dispensing hydrochloric acid as stop solution (change from blue to yellow). The intensity of the color formed is proportional to the concentration of TSH in the sample. The Optical Density (OD) of the color solution is measured with a microtiter plate reader at 450 nm. A calibrator curve is constructed by plotting OD values against concentrations of calibrators, and concentrations of unknown samples are determined using this calibrator curve.