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The FSH ELISA Test is an immunoassay designed for the quantitative determination of follicle-stimulating hormone concentrations in serum or plasma.

Human Follicle Stimulating Hormone [FSH] ELISA Kit


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Background

Follicle-Stimulating Hormone (FSH) and Luteinizing Hormone (LH) are intimately involved in the control of the growth and reproductive activities of the gonadal tissues, which synthesize and secrete male and female sex hormones. The levels of circulating FSH and LH are controlled by these sex hormones through a negative feedback relationship. FSH is a glycoprotein secreted by the basophilic cells of the anterior pituitary. Gonadotropin-release hormone (GnRH), produced in the hypothalamus, controls the release of FSH from the anterior pituitary. Like other glycoproteins, such as LH, TSH, and hCG, FSH consists of subunits designated as alpha and beta. Hormones of this type have alpha subunits that are very similar structurally, therefore the biological and immunological properties of each are dependent on the unique beta subunit. In the female, FSH stimulates the growth and maturation of ovarian follicles by acting directly on the receptors located on the grannulosa cells; follicular steroidogenesis is promoted and LH production is stimulated. The LH produced then binds to the theca cells and stimulates steroidogenesis. Increased intraovarian Estradiol production occurs as follicular maturation advances, thereupon stimulating increased FSH receptor activity and FSH follicular binding. FSH, LH, and Estradiol are therefore intimately related in supporting ovarian recruitment and maturation of theovum in female.

Test Principle

The FSH ELISA Test Kit is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a rabbit polyclonal anti-Primate FSH antibody for solid phase (microtiter wells) immobilization and mouse anti-Primate FSH antibody in coupled to enzyme (horseradish peroxides) conjugate solution. The test sample is allowed to react simultaneously with the antibodies, resulting in FSH molecules being sandwiched between the solid phase and enzyme-linked antibodies. After a 1hour incubation period at 37C, the wells are washed with buffer to remove unbound labeled antibodies. A solution of 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 2N HCl, and the absorbency is measured spectrophotometrically at 450nm. The intensity of the color formed is proportional to the amount of enzyme present and is inversely related to the amount of unlabeled FSH in the sample. By reference to a series of FSH standards assayed in the same way, the concentration of FSH in the unknown sample is quantified.

Feature and Advantages

< Examples of ETI Primate FSH Standard Curve >
A typical FSH ELISA standard curve run as a quality control of each lot is given below:

Primate FSH concentration ng/mL Absorbency 450nm
0.0 ng/ml 0.08
1 ng/ml 0.17
2.5 ng/ml 0.36
5.0 ng/ml 0.65
10 ng/ml 1.15
25 ng/ml 2.85

<ELISA Performance Characters>
Precision : Inter and Intra assay variation (CV) was determined from three different pooled serum samples in three different experiments.

Inter-assay variation Set1: CV= 8.2% (N=10) Set2: CV= 6.6 % (N=10) Set3: CV= 6.4 % (N=10)
Intra-assay variation Set1: CV= 7.8% (N=10) Set2: CV= 7.2 % (N=10) Set3: CV= 6.2 % (N=10)

< Sensitivity and Specificity >
The sensitivity of the assay is 0.5 ng/mL and each laboratory should establish its own base levels based on the species and physiological situation.
In ETI Primate FSH ELISA assay, the following materials have the following cross-rectivity : Primate FSH 100%, TSH < 0.08%, HCG < 0.01%, LH < 0.05% GH <0.001%.

< References >
1. Knobil, E. The neuroendocrine control of the menstrual cycle, Rec. Prog. Horm. Res. 36:52-88; 1980
2. Harris, G.W. and Naftolinf. The hypothalamus and control of ovulation. Brit. Med. Bullet. 26: 1-9; 1970
3. Shome, B. and Parlow, A.F. J. Clin. Endocrinol. Metabl. 39:199-205; 1974
4. Uotila, M.; Ruoslahti, E. and Engvall, E. J. Immunol. Methods. 42: 11-15; 1981
5. Murphy BD, et al: J Repod Fertil Suppl.1993;47:181-8
6. Valtonen M, et al: J Reprod Fertil Suppl.1993;47:133-7
7. Marshall J.C. Clinic in Endocrinol. Metab. 1975; 4:454
8. Cohen K.L. Mtabolism 1977; 26:1165
9. Rebar R.W., Erickson G.F. and Yen S.S.C. Fertil. Steril. 1982; 37:35
10. Abraham G.E. Ed. Radioassay Systems in Clinic. Endocrinol. Marecel Dekker, Inc., New York (1981)

Kit component

1. Antibody-coated microtiter wells, 96-well plate
2. Reference Standard, 0.8ml (0, 1.0, 2.5, 5.0, 10, 25, ng/mL)
3. Enzyme Conjugate Reagent, 12 mL
4. TMB Color Reagent, 12 mL
5. 20X Wash buffer, 20 mL
6. Stop solution (2N HCl), 6mL
7. Instructions

Product List

Product Name Cat# Quantity Price

FSH ELISA, Human DataSheet

ENC-ERKP6007

1 KIT ¥230,000
$3067

Note : These products cannot be sold to USA.

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