Thyroid Stimulating Hormone, Human (TSH) BioAssay(TM) ELISA Kit

Sample Type:\nSerum\n\nIntended Use:\nFor the quantitative determination of thyroid stimulating hormone (TSH) concentration in human serum. The assay is useful in the diagnosis of thyroid or pituitary disorders.\n\nIntroduction:\nThe determination of serum or plasma levels of thyroid stimulating hormone (TSH or thyrotropin) is recognized as a sensitive method in the diagnosis of primary and secondary hypothyroidism (1). TSH is secreted by the anterior lobe of the pituitary gland and induces the production and release of thyroxine and triiodothyronine from the thyroid gland (2). It is a glycoprotein with a molecular weight of approximately 28,000D, consisting of two chemically different subunits, alpha and beta (3). Although the concentration of TSH in the blood is extremely low, it is essential for the maintenance of normal thyroid function. The release of TSH is regulated by a TSH-releasing hormone (TRH) produced by the hypothalamus. The levels of TSH and TRH are inversely related to the level of thyroid hormone. When there is a high level of thyroid hormone in the blood, less TRH is released by the hypothalamus, so less TSH is secreted by the pituitary. The opposite action will occur when there is decreased thyroid hormone in the blood. This process is known as a negative feedback mechanism and is responsible for maintaining the proper blood levels of these hormones (4,5). TSH and the pituitary glycoproteins: luteinizing hormone (LH), follicle-stimulating hormone (FSH), and human chorionic gonadotropin (hCG), have identical alpha chains. The beta chains are distinct but do contain regions with identical amino acid sequences. These regions of homology can cause considerable cross-reactivity with some polyclonal TSH antisera. The use of a monoclonal antibody in this TSH ELISA test eliminates such crossreactivity, which could result in falsely elevated TSH values in either menopausal or pregnant females -- a population whose evaluation of thyroid status is clinically significant (6,7,8).\n\nTest Principle:\nThe TSH ELISA test is based on the principle of a solid phase enzyme-linked immunosorbent assay (9,10). The assay system utilizes a unique monoclonal antibody directed against a distinct antigenic determinant on the intact TSH molecule. Mouse monoclonal anti-TSH antibody is used for solid phase mobilization (on the microtiter wells). A goat anti-TSH antibody is in the antibodyenzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react simultaneously with the two antibodies, resulting in the TSH molecules being sandwiched between the solid phase and enzyme-linked antibodies. After a 60-minute incubation at room temperature, the wells are washed with water to remove unbound labeled antibodies. A solution of TMB Reagent is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of Stop Solution, changing the color to yellow. The concentration of TSH is directly proportional to the color intensity of the tests. Absorbance is measured spectrophotometrically at 450 nm.\n\nKit Components:\nMurine Monoclonal Anti-TSH-coated microtiter wells.\nStandard (0IU/ml), 1x1 vial\nStandard, (0.5IU/ml), 1x1 vial\nStandard, (2IU/ml), 1x1 vial\nStandard, (5IU/ml), 1x 1vial\nStandard, (10IU/ml), 1x 1vial\nStandard, (25IU/ml), 1x1vial \nEnzyme Conjugate Reagent, 13ml\nTMB Reagent (One-Step), 11ml\nStop Solution (1N HCl), 11ml.\n\nStorage and Stability:\nStore all components at 4 degrees C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

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SPECIFICATIONS

Catalog Number

T5400-45H

Size

1Kit

References

1 Burger, H. G., Patel, Y. C., Thyrotropin releasing hor Clinic. Endocrinol. and Metab., 6, 831-00(1977). h n data and stan curve. 2 Ezrin, C., The Thyroid, S. C. Werner and S. H. lngbar (eds.), Harper and Row, Hagerstown, MD, 9, 174-178 (1978). 3 Pierce, J. G., Endocrinology, 89, 1331-1344 (1971). 4 Berger, S. and Quinn, J. L., Fund. Clin. Chem., N. W. Tietz (ed.), W. B. Saunders Co., Phila., PA 14, 824-848 (1976). 5 Utiger, R. D., The Thyroid, S.C. Werner and S. H. Ingbar (eds.), Harper and Row, Hagerstown, MD, 9, 196-205 (1978). 6 Soos, M. and Siddle, K., J. Immun. Methods, 51, 57-68 (1982). 7 Wada, H. G., Danisch, R. J., Baxter, S. R., et al, Clin. Chem.,28, 1862-1866 (1982). 8 Snyder, P. J. and Utiger, R. D., J. Clin. Endocrinol. Metab., 34, . J. (eds.), Academic Press, New York, 419- 492(1980). 10 Uotila, M., Ruoslahti, E. and Engvall, E., J. lmmunol. Metho (1972). 9 Engall, E., Methods in Enzymology, Volume 70, Van Vunakis, H. and Langone, J 11-15 (1981).