The specificity of DHT ELISA kit was determined by measuring interference from high concentrations of the following; DHT (100%), Testosterone (8.7%), 5-beta-DHT (2%), Androstenedione (0.2%), Dehydroepiandrosterone sulfate, 17-beta-estradiol, Estriol, Estrone, Progesterone, 17-OH-Progesterone, Cortical pregnenolone were all <0.01%. DHT kit has been designed and tested for human serum samples. It may be optimized for other biological fluids. It has not been tested in animals (rat, mouse, etc). Since the steroid hormone is the same in all species, this kit should work in most species as long as the sample concentration is within the range of this kit.
DHT ELISA kit is based on simultaneous binding of DHT from samples and DHT-HRP conjugate to anti-DHT immobilized on the microtiter well plates. Therefore, free DHT and enzyme-bound DHT compete for limited antibody bound to the pale. In the absence of free DHT, there is a maximum amount of DHT-enzyme bound to the plate. At the end it will produce the maximum color. As the amount of free DHT (samples or std) increases, it reduces the amount of enzyme bound to the plate. The color (blue) produced is inversely proportional to the concentration of DHT in the sample. A std. curve is constructed by generating color from no DHT (std. A, 0pg/ml DHT) to the highest std (Std E, 2500pg/ml DHT). Adding stopping solution terminates the reaction. Absorbance is then measured on a microtiter well ELISA reader at 450nm. and the concentration of DHT in samples and control is read off the standard curve.
Dihydrotestosterone (DHT) (Full name, 5a-Dihydrotestosterone, abbreviating to 5a-DHT, androstanolone) is a biologically active metabolite of the hormone testosterone, formed primarily in the prostate gland, testes, hair follicles, and adrenal glands by the enzyme 5a-reductase by means of reducing the 4,5 double-bond. Dihydrotestosterone belongs to the class of compounds called androgens, also commonly called androgenic hormones or testoids. Androgens are part of the biology of gender by stimulating and controlling the development and maintenance of masculine characteristics. DHT is 3 times more potent than testosterone; testosterone is 5-10 times more potent than adrenal androgens. While DHT is best known for its roles in causing male pattern hair loss and prostate problems, it is crucial to virilization and is necessary to mitigate estrogen's effects in men. DHT is an important contributor to other characteristics generally attributed to males, including facial and body hair growth, and deepening of the voice. Unlike other androgens such as testosterone, DHT cannot be converted by the enzyme aromatase to estradiol. It, therefore, is frequently used in research settings to distinguish between effects of testosterone caused by binding to the androgen receptor, and those caused by testosterone's conversion to estradiol and subsequent binding to estrogen receptors.
Serum. DHT ELISA kit has been used in mouse/rat serum and plasma and culture medium (see published references). Other fluids may be adapted
D8036-01A: Microtiter Plate 1x96 wells
D8036-01B: Standard 0pg/ml, 1x10 ml
D8036-01C: Standard 25pg/ml 1x 600ul
D8036-01D: Standard 100pg/ml 1x 600ul
D8036-01E: Standard 500pg/ml 1x 600ul
D8036-01F: Standard 1000pg/ml 1x 600ul
D8036-01G: Standard 2500pg/ml 1x 600ul
D8036-01H: DHT Low Control 1x 600ul
D8036-01J: Assay Buffer 1x15 ml
D8036-01K: DHT (HRP), 100X 1x200ul
D8036-01L: TMB Substrate 1x16ml
D8036-01M: Wash Buffer, 10X 1x50ml
D8036-01N: Stop Solution (1N H2SO4) 1x6ml
D8036-01P: DHT High Control 1x 600ul
Storage and Stability:
Store at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.