Instant ELISA Kit for Dihydrotestosterone (DHT)

5α-DHT; 5α-Dihydrotestosterone; Androstanolone; Stanolone

Specificity

This assay has high sensitivity and excellent specificity for detection of Instant Dihydrotestosterone (DHT).
No significant cross-reactivity or interference between Instant Dihydrotestosterone (DHT) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Instant Dihydrotestosterone (DHT) and the recovery rates were calculated by comparing the measured value to the expected amount of Instant Dihydrotestosterone (DHT) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 101
EDTA plasma(n=5) 83-101 92
heparin plasma(n=5) 78-98 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Instant Dihydrotestosterone (DHT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Instant Dihydrotestosterone (DHT) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Instant Dihydrotestosterone (DHT) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-103% 79-93% 95-105% 80-101%
EDTA plasma(n=5) 82-103% 93-101% 91-104% 82-93%
heparin plasma(n=5) 86-101% 87-95% 94-102% 91-98%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 5 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 30 minutes at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 30 minutes at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 10 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Internation Scholarly Research Network Herbicide Metolachlor Causes Changes in Reproductive Endocrinology of Male Wistar Rats Isrn: 130846
Journal of American Science Metabolic Changes and Hormonal Disturbances in Polycystic Ovarian Syndrome Rats and the Amelioration Effects of Metformin and/or Cinnamon Extraction Jofamericanscience: Source
Journal of Ethnopharmacology Treatment of benign prostatic hyperplasia with Croton membranaceus in an experimental animal model Pubmed:25256687
Toxicol Ind Health Oral administration of low-dose bisphenol A promotes proliferation of ventral prostate and upregulates prostaglandin D2 PubMed: 26088557
PPAR Research Testosterone Replacement Modulates Cardiac Metabolic Remodeling after Myocardial Infarction by Upregulating PPARα pubmed:27413362
Neurochemical Research Dihydrotestosterone Treatment Accelerates Autograft Reversal Sciatic Nerve Regeneration in Rats Pubmed:29340845
Journal of Endocrinology Uterine progesterone signaling is a target for metformin therapy in polycystic ovary syndrome Pubmed: 29535146
Journal of Cellular Physiology Dihydrotestosterone synthesis in the sheep corpus luteum and its potential mechanism in luteal regression Pubmed: 30671954
PHYTOTHERAPY RESEARCH Plant cell culture extract of Cirsium eriophorum with skin pore refiner activity by modulating sebum production and inflammatory response Pubmed: 32816329
Curr Oncol Plasma Aromatase Activity Index, Gonadotropins and Estrone Are Associated with Frailty Syndrome in Post-Menopausal Women with Breast Cancer Pubmed:35323344
J Steroid Biochem Mol Biol Syce1 and Syce3 regulate testosterone and dihydrotestosterone synthesis via steroidogenic pathways in mouse Sertoli and Leydig cells Pubmed:35697131
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