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|Method & Line||Sample & Target||Product||Package Info|
|MicroELISA||Plasma,Serum||EIAgen Free Testosterone Kit||Tests per Package: 96|
|EIAgen||The EIAgen Free Testosterone Kit has been designed for the quantitative determination of Free Testosterone Concentration in Human Serum or Plasma.||Code: LI10042F1||Package: 1 Microplate|
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The EIAgen Free Testosterone Kit has been designed for the quantitative determination of Free Testosterone Concentration in Human Serum or Plasma.
Testosterone, (17b-Hydroxy-4-androstene-3-one), a C19 steroid, is the most potent naturally secreted androgen1. In normal post pubertal males, testosterone is secreted primarily by the testes with only a small amount derived from peripheral conversion of 4-Androstene-3, 17-dione (ASD)2. In adult women, it has been estimated that over 50% of serum testosterone is derived from peripheral conversion of ASD secreted by the adrenal and ovary, with the remainder from direct secretion of testosterone by these glands.
In the male, testosterone is mainly synthesized in the interstitial Leydig cells and the testis, and is regulated by the interstitial cell stimulating hormone (ICSH), or luteinizing hormone (LH) of the anterior pituitary (the female equivalent of ICSH)3. Testosterone is responsible for the development of secondary sex characteristics, such as the accessory sex organs, the prostate, seminal vesicles and the growth of facial, pubic and auxiliary hair. Testosterone measurements have been very helpful in evaluating hypogonadal states. Increased testosterone levels in males can be found in complete androgen resistance (testicular feminization). Common causes of decreased testosterone levels in males include: hypogonadism, orchidectomy, estrogen therapy, Klinefelter's syndrome, hypopituitarism, and hepatic cirrhosis2-4.
In the female, testosterone levels are normally found to be much lower than those encountered in the healthy male. Testosterone in the female comes from three sources. It is secreted in small quantities by both the adrenal glands and the ovaries, and in healthy women 50–60% of the daily testosterone production arises from peripheral metabolism of prohormone, chiefly androstenedione. Common causes of increased serum testosterone levels in females include polycystic ovaries (Stein-Leventhal syndrome), ovarian tumors, adrenal tumors and adrenal hyperplasia. Virilization in women is associated with the administration of androgens and endogenous overproduction of testosterone. There appears to be a correlation between serum testosterone levels and the degree of virilization in women, although approximately 25% of women with varying degrees of virilism have serum testosterone levels that fall within the female reference range.
The majority of testosterone is bound to transport proteins; weakly bound to albumin and cortisol binding protein (25-65% females - 45-85% males) and tightly bound to sex hormone-binding globulin (SHBG) (females 35-75% - males 14-50%)8. A small fraction exist as unbound or free testosterone, however this form is biologically active. Therefore, the free hormone concentration is a better indicator of biological activity than total testosterone.
The essential reagents required for a enzyme immunoassay include antibody, enzyme-antigen conjugate and native antigen.
Upon mixing biotinylated antibody, enzyme-antigen conjugate and a serum containing the native free antigen, a competition reaction results between the native free antigen and the enzyme-antigen conjugate for a limited number of antibody binding sites.
A simultaneous reaction between the biotin attached to the antibody and the streptavidin immobilized on the microwell occurs. This effects the separation of the antibody bound fraction after decantation or aspiration.
The enzyme activity in the antibody bound fraction is inversely proportional to the native antigen concentration. By utilizing several different serum references of known antigen concentration, a dose response curve can be generated from which the antigen concentration of an unknown can be ascertained.
The kit contains reagents for 96 tests (code LI10042F1).
Bring reagents to room temperature before use.
Free Testosterone Biotin
Wash Buffer Conc. 50X
Number of tests