Progesterone ELISA

Enzyme Immunoassay for the Quantitative Determination of Progesterone Concentration in Human Serum

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES

SUMMARY
Progesterone is a C21 steroid which is synthesized from both tissue and circulating cholesterol. Cholesterol is transformed to pregnenolone which is then converted via a combined dehydrogenase and isomerase to progesterone. The principle production sites are the adrenals and ovaries and the placenta during pregnancy. The majority of this steroid is metabolized in the liver to pregnanediol and conjugated as a glucuronide prior to excretion by the kidneys.

Progesterone exhibits a wide variety of end organ effects. The primary role of progesterone is exhibited by the reproductive organs. In males, progesterone is a necessary intermediate for the production of corticosteroids and androgens. In females, progesterone remains relatively constant throughout the follicular phase of the menstrual cycle. The concentration then increases rapidly following ovulation and remains elevated for 4-6 days and decreases to the initial level 24 hours before the onset of menstruation. In pregnancy, placental progesterone production rises steadily to levels of 10 to 20 times those of the luteal phase peak.

Progesterone measurements are thus performed to determine ovulation as well as to characterize luteal phase defects. Monitoring of progesterone therapy and early stage pregnancy evaluations comprise the remaining uses of progesterone assays.

United Immunoassay Progesterone EIA kits are designed for the measurement of total progesterone in human serum or plasma.

PRINCIPLE OF THE TEST
The wells are washed with water to remove unbound-labeled antibodies. A solution of TMB is added and incubated for 20 minutes at room temperature, resulting in the development of a blue color. The color development is stopped with the addition of 1N HCl, changing the color to yellow. The concentration of Beta2-MG is directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450nm.

EXAMPLE OF STANDARD CURVE
Results of a typical standard run with optical density readings at 450 nm shown in the Y axis against Progesterone concentrations shown in the X axis. This standard curve is for the purpose of illustration only, and should not be used to calculate unknowns. Each user should obtain his or her own data and standard curve.