Pregnenolone ELISA Kit


Human pregnenolone ELISA kit is a reliable quantitative procedure for measuring pregnenolone (3β-hydroxypregn-5-en-20-one) in human serum. This assay has a minimum sensitivity detection limit of 0.05 ng/ml.


Pregnenolone (which is also referred to as 3β-hydroxypregn-5-en-20-one) is a one of the most common precursor the majority of adrenal and gonadal steroids. It was the first steroid hormone which was derived from cholesterol from the pathway of steroidogenesis. Human pregnenolone is produced within the mitochondria as a result of the C-20 side chain of cholesterol being cleaved by the P-450SCC enzyme. After it has been produced, it can be utilised by two pathways of steroidogenesis. One is through enzymatic action of 3β-hydroxysteroid dehydrogenase where it is converted to progesterone and the other via enzymatic action of 17α-hydroxylase where it is converted to 17-OH pregnenolone.

Studies of pregnenolone concentrations in relation to sex and age differences has been found to indicate that maximum levels occur in men at 17 ages and in women at age 16. Whilst the minimum levels occur in men at age 28 and in women at age 37. In general women have been found to have higher levels when compared to men, also higher levels are discovered in women that have idiopathic hirsutism. Elevated human pregnenolone concentrations occur in forms of congenital adrenal hyperplasia (CAH), this is caused by a deficiency in either 17α-hydroxylase or 3β-hydroxysteroid dehydrogenase.

There are many areas of pregnenolone physiology which is still not investigated, however, many current publications indicate that the measurement of human pregnenolone in serum could be helpful in studying its metabolite that have been reported to have a number of effects on the brain and central nervous system.


All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • Microtiterwells: Break apart strips that are coated with anti-pregnenolone antibody (polyclonal).
  • Calibrators 0-6: Concentrations 0, 0.1, 0.4, 1.6, 6.4, 25.6 ng/ml.
  • Control: Contains anti pregnenolone in a human serum matrix.
  • Pregnenolone Biotin Enzyme Conjugate (50x).
  • Streptavidin-Horseradish Peroxide (HRP) Conjugate (50x).
  • Biotin Conjugate Buffer.
  • HRP Conjugate Buffer.
  • Wash Buffer (10x Concentrated).
  • TMB Substrate.
  • Stopping Solution.


The minimum sensitivity detection limit of 3β-hydroxypregn-5-en-20-one using current human pregnenolone ELISA kit was approximately 0.05 ng/ml. The dynamic range for this assay is 0.1 – 25.0 ng/ml.


– Pregnenolone: ELISA
– Sensitivity: 0.054 ng/ml
– Specificity (Cross Reactivity): Pregnenolone (100%), Progesterone (6.0%), Dehydroisoandrosterone (5.2%), 5α-Androstandiol (4.7%), Epiandrosterone (1.0%), Pregnenolone Sulfate (0.4%), Androstandione (0.3%), 5α-Androsterone (0.3%), DHEAS (0.2%), Etiocholanolone (0.1%), Aldosterone, Androstenedione, Cholesterol, Corticosterone, 5α-DHT, 17β-Estradiol, Estriol, Testosterone (< 0.1%).
– Intra-Assay Variability: 8.7 – 9.8%
– Inter-Assay Variability: 8.4 – 11.7%
– Recovery: 101.5 – 118.3%
– Linearity: 82.9 – 102.2%
– Expected Normal Values: Males (0.38 – 3.5 ng/ml), Females (0.31 – 3.8 ng/ml).


  1. Neurosteroid regulation of central nervous system development. Pharmacol Ther. (2007) 116 (1): 107-24. Review. Mellon S.H.
  2. Progesterone synthesis by the human placenta. Placenta. (2005) 26 (4): 273-81. Review. Tuckey RC.
  3. Role of pregnenolone, dehydroepiandrosterone and their sulfate esters on learning and memory in cognitive aging. Brain Res Brain Res Rev. (2001) 37 (1-3): 301-12. Review. Vallée M., et al.
  4. Neurosteroids and potential therapeutics: Focus on pregnenolone. J Steroid Biochem Mol Biol. (2016) 160: 78-87. Review. Vallée M.


  • Full Name: Pregnenolone ELISA Kit
  • Reactivity: Human
  • Sample Type: Serum
  • Sensitivity: 0.05 ng/ml



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