Prolactin, 1 x 96 Assays

FOR RESEARCH USE ONLY.

Product Name: Prolactin

Sample Type: Serum

# of Samples: 1X96 Assays

Intended Use: The GenWay Biotech Prolactin ELISA is intended for the quantitative determination of prolactin in human serum.

Introduction: Human prolactin (lactogenic hormone) is secreted from the anterior pituitary gland in both men and women^1. Human prolactin is a single chain polypeptide hormone with a molecular weight of approximately 23,000 daltons.^2 The release and synthesis of prolactin is under neuroendocrinal control, primarily through Prolactin Releasing Factor and Prolactin Inhibiting Factor.^3
Women normally have slightly higher basal prolactin levels than men; apparently there is an estrogen-related rise at puberty and a corresponding decrease at menopause. The primary functions of prolactin are to initiate breast development and to maintain lactation. Prolactin also suppresses gonadal function.^4,5
During pregnancy, prolactin levels increase progressively to between 10 and 20 times of normal values, declining to non-pregnant levels by 3-4 weeks post-partum.^4 Breast-feeding mothers maintain high levels of prolactin, and it may take several months for serum concentrations to return to non-pregnant levels.^3,4
Microadenomas (small pituitary tumors) may cause hyperprolactinemia, which is sometimes associated with male impotence.6 High prolactin levels are commonly associated with galactorrhea and amenorrhea.^5
Prolactin concentrations have been shown to be increased by estrogens, thyrotropin-releasing hormone (TRH), and several drugs affecting dopaminergic mechanisms.^7,8,9,10 Prolactin levels are elevated in renal disease and hypothyroidism, and in some situations of stress, exercise, and hypoglycemia. Additionally, the release of prolactin is episodic and demonstrates diurnal variation.^5 Mildly elevated prolactin concentrations should be evaluated taking these considerations into account. Prolactin concentrations may also be increased by drugs such as chloropromazine and reserpine, and may be lowered by bromocyptine and L-dopa.^4
The GenWay Biotech Prolactin Enzyme Immunoassay provides a rapid, sensitive, and reliable assay for the measurement of prolactin. The antibodies developed for the test will determine a minimal concentration of human prolactin of 2 ng/ml. There is no cross-reactivity with hCG, TSH, LH, FSH, or hGH.

Principles of the assay: The GenWay Biotech Prolactin Quantitative Test is based on the principle of a solid phase enzyme-linked immunosorbent assay.^11,12 The assay system utilized sheep anti-prolactin for solid phase (microtiter wells) immobilization and mouse monoclonal anti-prolactin in the antibody-enzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react simultaneously with the antibodies, resulting in the prolactin molecules being sandwiched between the solid phase and enzyme-linked antibodies. After a 45 minute incubation at room temperature, the wells are washed with water to remove unbound labeled antibodies. A solution of Tetramethylbenzidine (TMB) is added and incubated for 20 minutes, resulting in the development of a blue color. The color development is stopped with the addition of 1N HCl, and the resulting yellow color is measured spectrophotometrically at 450 nm. The concentration of prolactin is directly proportional to the color intensity of the test sample.

Storage and Stability: 1. Store the unopened kit at 2-8C upon receipt and when it is not in use, until the expiration shown on the kit label. Refer to the package label for the expiration date.
2. The opened and used reagents are stable until the expiration date if stored properly at 2-8C.
3. Keep microtiter plate in a sealed bag with desiccant to minimize exposure to damp air.

Limitation of the Test: 1. Reliable and reproducible results will be obtained when the assay procedure is carried out with a complete understanding of the package insert instructions and with adherence to good laboratory practice.
2. Serum samples demonstrating gross lipemia, gross hemolysis, or turbidity should not be used with this test.
3. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
4. Pregnancy, estrogen treatment, renal disease, and hypothyroidism may affect prolactin levels.