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Specifications
- Assay Type Competitive ELISA
- Sample Types Extracted Serum, Extracted Plasma, Urine, Fecal Extracts, Tissue Culture Media
- Sensitivity 20.3 pg/mL
- Species 17HO-P is identical across species
- Assay Duration 1.5 Hours
- Samples/Plate 40 in Duplicate
- Readout Colorimetric, 450 nm
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Standard Curve
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Description
Assay Principle:
The 17-Hydroxyprogesterone ELISA Kit quantitatively measures 17-Hydroxyprogesterone (17HO-P) in extracted serum, extracted plasma, urine, fecal extracts, and tissue culture media. The 17-Hydroxyprogesterone ELISA Kit is a competitive ELISA with a run time of 1.5 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided 17HO-P standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our donkey anti-sheep IgG antibody. Add the 17HO-P peroxidase conjugate and the 17HO-P polyclonal sheep antibody. Then incubate the mixture covered at room temperature, shaking for 1 hour. The immunological reaction occurs between the anti-17HO-P antibody, the 17HO-P antigen in the sample or standard, and the 17HO-P-peroxidase conjugate. As the 17HO-P concentration in the sample increases, the bound 17HO-P-peroxidase conjugate decreases, causing a decrease in signal and vice versa.
After the 1-hour incubation, wash away the excess 17HO-P-peroxidase conjugate and add the TMB substrate. The TMB substrate reacts with the bound 17HO-P-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the 17HO-P concentration in the samples.
Background:
17-Hydroxyprogesterone is a steroid hormone belonging to the androgen group, found in mammals, reptiles, birds, and other vertebrates. Pfiffner and North first isolated it in 1940. It is primarily produced in the adrenal glands and the ovary’s corpus luteum. It is hydroxylated at the 11 and 21 positions to produce cortisol. A deficiency of either 11- or 21-hydroxylase results in decreased cortisol synthesis, and feedback inhibition of adrenocorticotropic hormone (ACTH) secretion is lost. Consequently, the increased pituitary release of ACTH will increase the production of 17HO-P. However, if 17-alpha-hydroxylase or 3β-hydroxysteroid dehydrogenase type 2 is deficient, then 17HO-P levels will be low with either a possible increase in progesterone or pregnenolone, respectively. Normal levels are 3-90 ng/dL in children. In women, normal levels are 20-100 ng/dL before ovulation and 100-500 ng/dL during the luteal phase.
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Structure