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Research Areas
Documentation
Specifications
- Assay Type Competitive ELISA
- Sample Types Urine, Extracted Serum, Extracted Plasma (EDTA and Heparin), Dried Feces, Tissue Culture Media
- Sensitivity 129.7 pg/mL
- Species Allopregnanolone is identical across species
- Assay Duration 2.5 Hours or Overnight
- Samples/Plate 39 in Duplicate
- Readout Colorimetric, 450 nm
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Standard Curve
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Description
Assay Principle:
The Allopregnanolone ELISA Kit quantitatively measures allopregnanolone in urine, extracted serum, extracted plasma (EDTA and Heparin), dried feces, and tissue culture media. The Allopregnanolone ELISA Kit is a competitive ELISA with a run time of 2.5 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided allopregnanolone standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our goat anti-mouse IgG antibody. Add the allopregnanolone peroxidase conjugate and the allopregnanolone mouse monoclonal antibody. Then incubate the mixture covered at room temperature, shaking for 2 hours. The immunological reaction occurs between the anti-allopregnanolone antibody, the allopregnanolone antigen in the sample or standard, and the allopregnanolone conjugate. As the allopregnanolone concentration in the sample increases, the bound allopregnanolone-peroxidase conjugate decreases, causing a decrease in signal and vice versa.
After the 2-hour incubation, wash away the excess allopregnanolone-peroxidase conjugate and add the TMB substrate. The TMB substrate reacts with the bound allopregnanolone-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the allopregnanolone concentration in the samples.
Background:
Allopregnanolone (3α-hydroxy-5α-pregnan-20-one, THP, THPROG) is a prototypic neurosteroid in the blood and the brain. It is a metabolite of progesterone and a potent modulator of GABAA receptors. Allopregnanolone has pharmacological properties, including anxiolytic and anticonvulsant activity. The biosynthesis of allopregnanolone involves the conversion of progesterone into 5α-dihydro progesterone by the enzyme 5α-reductase type I. Subsequently, 3α-hydroxysteroid oxidoreductase isoenzymes convert this intermediate into allopregnanolone. Anxiety and depression are common side effects of 5α-reductase inhibitors such as finasteride and dutasteride. The prevention of the endogenous production of allopregnanolone by these inhibitors may be responsible for these side effects. Allopregnanolone aids neurogenesis and can reverse proliferative neuron deficits and cognitive deficits in mouse models of Alzheimer’s disease.