Nori Canine TSH ELISA Kit Summary

Alternative names for TSH: Thyroid-stimulating hormone, thyrotropin, thyrotropic hormone,

Alternative names for canine: Dog

CCP37964

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Thyroid-stimulating hormone (also known as thyrotropin, thyrotropic hormone, TSH. TSH is a pituitary hormone that stimulates the thyroid gland to produce thyroxine (T₄), and then triiodothyronine (T₃) which stimulates the metabolism of almost every tissue in the body. TSH is a glycoprotein and consists of two subunits, the alpha and the beta subunit. It is a hormone synthesized and secreted by thyrotrope cells in the anterior pituitary gland, which regulates the endocrine function of the thyroid. TSH (with a half life of about an hour) stimulates the thyroid gland to secrete the hormone thyroxine (T₄), which has only a slight effect on metabolism. T₄ is converted to triiodothyronine (T₃), which is the active hormone that stimulates metabolism. About 80% of this conversion is in the liver and other organs, and 20% in the thyroid itself. TSH is secreted throughout life but particularly reaches high levels during the periods of rapid growth and development. The hypothalamus, in the base of the brain, produces thyrotropin-releasing hormone (TRH). TRH stimulates the pituitary gland to produce TSH. Somatostatin is also produced by the hypothalamus, and has an opposite effect on the pituitary production of TSH, decreasing or inhibiting its release. The concentration of thyroid hormones (T₃ and T₄) in the blood regulates the pituitary release of TSH; when T₃ and T₄ concentrations are low, the production of TSH is increased, and, conversely, when T₃ and T₄ concentrations are high, TSH production is decreased. This is an example of a negative feedback loop.^[1] Any inappropriateness of measured values, for instance a low-normal TSH together with a low-normal T₄ may signal tertiary (central) disease and a TSH to TRH pathology. Elevated reverse T₃ (RT₃) together with low-normal TSH and low-normal T₃, T₄ values, which is regarded as indicative for euthyroid sick syndrome, may also have to be investigated for chronic subacute thyroiditis (SAT) with output of subpotent hormones. Absence of antibodies in patients with diagnoses of an autoimmune thyroid in their past would always be suspicious for development to SAT even in the presence of a normal TSH because there is no known recovery from autoimmunity. For clinical interpretation of laboratory results it is important to acknowledge that TSH is released in a pulsatile manner^[2][3] resulting in both circadian and ultradian rhythms of its serum concentrations.

References

1. Estrada JM, et al. (2014). Thyroid 24 (3): 411–23.
2. Brabant G, et al. (1990). J Clinical Endocrinology and Metabolism 70 (2): 403–9.
3. Samuels MH, et al. (1990). J Clinical Endocrinology and Metabolism 71 (2): 425–32.