The Rodent LH ELISA test is an immunoassay designed for the quantitative determination of luteinizing hormone (LH) in serum/plasma samples of Rodents and related species.

Rodent Luteinizing Hormone [LH] ELISA Kit


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Background

Luteinizing Hormone (LH) and Follicle-Stimulating Hormone (FSH) are intimately involved in the control of the growth andreproductive 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. LH is a glycoprotein secreted by the basophilic cells of the anterior pituitary. Gonadotropin-release hormone (GnRH or LHRH), produced in the hypothalamus, controls the release of LH and FSH from the anterior pituitary. Like other glycoproteins FSH, TSH, and hCG, LH consists of two subunits alpha and beta. All these hormones have structurally similar alpha subunit, unique beta subunit which determine the biological and immunological properties. In the male the hormone binds to Leydig cells and enhances the secretion of male hormone Testosterone. The LH binds to the theca cells and stimulates steroidogenesis in the ovary. 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 in females.

Test Principle

The LH ELISA Test Kit is based on the principle of a solid phase enzyme-linked immunosorbent assay. The assay system utilizes a polyclonal anti-LH antibody for solid phase (microtiter wells) immobilization and mouse anti-LH antibody in the antibody-enzyme (horseradish peroxidase) conjugate solution. The test sample is allowed to react simultaneously with the two antibodies, resulting in LH molecules being sandwiched between the solid phase and enzyme-linked antibodies. After a 2 hours incubation , the wells are washed with wash 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 directly related to the amount of unlabeled LH in the sample. By reference to a series of LH standards assayed in the same way, the concentration of LH in the unknown sample is quantified.

Feature and Advantages

<Quality Control Data>
It is highly recommended that each laboratory must establish their own internal controls and normal reference values for desired age, sex and physiological parameters.
A typical standard curve (illustration only) for rat/mouse LH is given below:

Standard ng/mL OD at 450nm
0 0.05
1 0.31
5 0.62
10 0.95
25 1.87
50 2.6

<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= 4.3% (N=10) Set2: CV= 5.8 % (N=10) Set3: CV= 5.4 % (N=10)
Intra-assay variation Set1: CV= 4.9% (N=10) Set2: CV= 5.4 % (N=10) Set3: CV= 5.9 % (N=10)

< Sensitivity >
The lowest level detectable in this assay is 0.5 ng/mL of serum or plasma

< Specificity >
The rat LH ELISA system utilizes monoclonal antibody and high affinity polyclonal antibody to LH. The cross reactivity to other pituitary gonadotropins (rat TSH, FSH is not detectable under the sensitivity of the assay system.

< 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. Goodman MF: Probl Vet Med. 1992, 4(3):433-44. Review.
6. Nishiyama T, et al.: J Am Anim Hosp Assoc. 1999, 35(4): 348-52
7. Chiba K, et al: Endocr J.1997, 44 (2): 205-18
8. Murphy BD, et al: J Repod Fertil Suppl.1993;47:181-8
9. Valtonen M, et al: J Reprod Fertil Suppl.1993;47:133-7
Revised 112004

Kit component

1. Antibody-coated microtiter wells, 96-well plate
2. Enzyme Conjugate Reagent, 12 mL
3. TMB Color Reagent (ready to use) , 12 mL
4. 20X Wash buffer, 20 mL
5. Stop solution (2N HCl), 6mL
6. Sample Diluent, 20 mL
7. Reference Standard/Ready to use/0.5mL/Vial
(0, 1, 2.5, 5, 10, 25, 50 ng/mL, store frozen –20C)
8. Instructions

Product List

Product Name Cat# Quantity Price

LH ELISA, Rodent DataSheet

ENC-ERKR7010

1 KIT ¥125,000
$1667

Note : These products cannot be sold to USA.

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