ELISA Kit for Androstenedione (ASD)

4-Androstenedione; 4-Androstene-3,17-Dione


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


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

Matrix Recovery range (%) Average(%)
serum(n=5) 88-102 98
EDTA plasma(n=5) 83-90 86
heparin plasma(n=5) 81-95 90


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


The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Androstenedione (ASD) 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) 87-96% 96-104% 96-103% 87-101%
EDTA plasma(n=5) 84-101% 79-101% 95-105% 94-102%
heparin plasma(n=5) 99-105% 85-92% 78-105% 79-101%


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.



Magazine Citations
Plos One Association between Polycystic Ovary Syndrome and Cavia (Guinea pig )t Microbiota Pubmed:27093642
Scientific Reports Molecular characterization of insulin resistance and glycolytic metabolism in the rat uterus pubmed:27461373
33 Metformin Ameliorates Uterine Defects in a Rat Model of Polycystic Ovary Syndrome. pubmed:28336389
Journal of Endocrinology Uterine progesterone signaling is a target for metformin therapy in polycystic ovary syndrome Pubmed: 29535146
International Journal of Biological Sciences Melatonin Stimulates STAR Expression and Progesterone Production via Activation of the PI3K/AKT Pathway in Bovine Theca Cells
Molecular reproduction and development Evidence that downregulation of Wilms' tumor 1 (WT1) is involved in cortical stromal cell differentiation into theca cells in adult bovine ovaries Pubmed: 31490589
DOMESTIC ANIMAL ENDOCRINOLOGY Wilms' tumor (WT1)(+/-KTS) variants decreases the progesterone secretion of bovine ovarian theca cells Pubmed: 32739762
ANIMAL REPRODUCTION SCIENCE A study on steroidogenic elaborations of stroma and their regulation in response to ovarian hormones in goats 33845412
Reprod Domest Anim Extracellular Vesicles of Bovine Small Follicular Fluid Promote Ovarian Cortical Stromal Cell Proliferation and Steroidogenesis 34402549
Curr Oncol Plasma Aromatase Activity Index, Gonadotropins and Estrone Are Associated with Frailty Syndrome in Post-Menopausal Women with Breast Cancer Pubmed:35323344
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