Steroid 5 alpha-Reductase Rat Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
The local conversion of testosterone to the more potent androgen dihydrotestosterone (DHT) by 5a-reductase plays a role in the development of androgenetic alopecia and hirsutism. Two distinct isozymes are found in mice, rats, monkeys, and humans: Type I and II. Each of these isozymes is differentially expressed in tissues and developmental stages. In human, Type I 5a-reductase is predominant in the sebaceous glands of most regions of skin, including scalp and liver, and is responsible for approximately one-third of circulating DHT. Type II 5a-reductase is primarily found in prostate, seminal vesicles epididymides, and hair follicies as well as liver, and is responsible for two-thirds of circulating DHT. Inhibitors of 5a-reductase may be of benefit in the treatment of androgenetic alopecia.
Family:
Oxidoreductase
Sub Family:
Reductase
Organism:
Rat
Construct Details:
Endogenous
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
EIA
Measured Response:
Absorbance
Testing Information
Procedure Summary:
Rat liver steroid 5 alpha-reductase is used. Test compound and/or vehicle is preincubated with 20 μg/ml enzyme in modified phosphate buffer pH 6.5 for 15 minutes at 37⁰C. The reaction is initiated by addition of 0.9 μM testosterone for another 30 minute incubation period and terminated by further addition of 1 N HCl. Determination of the amount of testosterone remained is measured spectrophotometrically by EIA kit. Compounds are screened at 10 μM.
Substrate:
0.9 µM Testosterone
Incubation:
30 min at 37°C
Control Inhibitor:
Dutasteride
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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