Androstenedione ELISA Kit (OKAU00063)

Catalog No: OKAU00063

ELISA Standard Curve

Product Info

• Application: Enzyme-linked immunosorbent assay
• ELISA Kit Linearity: Linearity was determined by taking two saliva samples diluted with Assay Buffer, one with a low androstenedione level and one with a higher level, and mixing them in the ratios given below. The measured concentrations were compared to the expected values based on the ratios used.

  High Saliva	Low Saliva	Expected Conc. (pg/mL)	Observed Conc. (pg/mL)	% Recovery
  80%	20%	167.1	168.4	100.8
  60%	40%	129.8	134.9	103.2
  40%	60%	92.6	90.1	97.3
  20%	80%	55.4	49.7	89.7
  			Mean Recovery	97.7%

• ELISA Kit Principle: The DetectX® Androstenedione Immunoassay Kit uses a specifically generated antibody to measure androstenedione in saliva or in extracted serum and plasma. This kit is not recommended for serum or plasma samples without extraction. The kit will also quantitatively measure androstenedione present in reconstituted buffer samples and tissue culture media samples. Please read the complete kit insert before performing this assay. An androstenedione standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture sheep antibodies. An androstenedione-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a polyclonal antibody to androstenedione to each well. After a two hour incubation the plate is washed and substrate added. The substrate reacts with the bound androstenedione-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. After making suitable correction for the dilution of the sample, the concentration of the androstenedione in the sample is calculated using software available with most plate readers.
• ELISA Kit Reproducibility: Intra Assay Precision Four saliva samples were diluted with Assay Buffer and run in replicates of 20 in an assay. The mean and precision of the calculated androstenedione concentrations were:

  Sample	Androstenedione Conc. (pg/mL)	%CV
  1	226.2	6.6
  2	116.0	9.4
  3	57.1	13.9
  4	31.7	9.2

  Inter Assay Precision Four saliva samples were diluted with Assay Buffer and run in duplicates in 19 assays run over multiple days by four operators. The mean and precision of the calculated androstenedione concentrations were:

  Sample	Androstenedione Conc. (pg/mL)	%CV
  1	232.4	9.7
  2	119.3	9.3
  3	58.9	7.4
  4	34.2	12.1

• ELISA Kit Component:
  • Coated Clear 96 Well Plates: 1 or 5 Each
  • Androstenedione Standard: 125 uL or 625 uL
  • DetectX® Androstenedione Antibody: 3 mL or 13 mL
  • DetectX® Androstenedione Conjugate: 3 mL or 13 mL
  • Assay Buffer Concentrate: 28 mL or 55 mL
  • Wash Buffer Concentrate: 30 mL or 125 mL
  • TMB Substrate: 11 mL or 55 mL
  • Stop Solution: 5 mL or 25 mL
  • Plate Sealer: 1 or 5 Each
• Additional Information: Background: Androstenedione is a steroid hormone also known as androst-4-ene-3,17-dione. It is synthesized from dehydroepiandrosterone (DHEA) or 17-hydroxyprogesterone. By itself, androstenedione is a weak androgen though it is a precursor for biosynthesis of stronger androgens, such as testosterone and estrogens. Therefore, androstenedione is classified as a controlled drug and a supplement to enhance athletic performance and body energy. In males, androgens are secreted by the testes and the adrenal cortex while, in females, they are secreted by the ovaries. The production of androstenedione by adrenal glands is partially regulated by andrenocorticotripic hormone (ACTH). The amount of androstenedione in body fluids fluctuates throughout the day, with levels measuring highest in the morning and lowest in the evening. The levels of androstenedione in serum also change with age and physiological conditions. During embryonic development, the levels increase with the highest values at birth and the levels then decrease during the first few years of development. At puberty, the levels rise again for a few years, then taper down during menopause in females. Generally, a higher level of androstenedione may provide androgenic risk to females indicating higher androgenic activity, hirsutism or polycystic ovarian syndromes. Due to androgenic and estrogenic function of this sex steroid, children and adolescents are highly sensitive to androstenedione exposure; thus, androstenedione is treated as a regulated substance by the DEA.
  Detection Limit: 1.48 pg/mL
  Cross Reactivity: The following cross reactants were tested in the assay and calculated at the 50% binding point.

  Steroid	Cross Reactivity (%)
  Androstenedione	100%
  Epiandrosterone	4.51%
  Dehydroepiandrosterone (DHEA)	0.98%
  Testosterone	0.48%
  17-Hydroxyprogesterone	0.09%
  Progesterone	0.03%
  Estrone	0.0%
  Estradiol	< 0.01%

• Reconstitution and Storage: 2°C to 8°C
• Sample Type: Saliva, Urine, Extracted Serum/Plasma
• Sensitivity: 2.30 pg/mL

Target Info

• Gene Full Name: Androstenedione