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ELISA Kit for Dihydrotestosterone (DHT)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 79-104 101
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 78-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Dihydrotestosterone (DHT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 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 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) 83-101% 98-105% 95-103% 78-101%
EDTA plasma(n=5) 82-93% 98-105% 98-105% 93-103%
heparin plasma(n=5) 80-88% 98-105% 80-102% 78-99%

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 2 Standard Diluent 1×20mL
Detection Reagent A 1 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

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