Assay Principle:

The DetectX® PGFM ELISA Kit quantitatively measures PGFM in various samples such as serum, plasma, fecal extracts, urine, and tissue culture media. This competitive ELISA has a run time of 1.5 hours and is designed for efficient and accurate measurement. For optimal assay performance, users should thoroughly read the kit insert before beginning the assay. The kit includes a PGFM standard to establish an accurate standard curve.

Protocol Summary:

• Place standards or diluted samples into the provided transparent microtiter plate coated with goat anti-rabbit IgG antibody.
• Add PGFM peroxidase conjugate and PGFM polyclonal rabbit antibody to initiate the immunological reaction.
• Incubate the mixture covered at room temperature with shaking for 1 hour, as the reaction inversely correlates with PGFM concentration in the sample.
• After incubation, wash away excess conjugate and add the TMB substrate, which reacts with the bound conjugate to produce a detectable signal.
• Use a plate reader to measure the signal at 450nm and calculate the PGFM concentration based on the standard curve.

Background:

Prostaglandin F2 (PGF2), predominantly produced in uterine and placental tissues, plays an important role in regulating reproductive and pregnancy-related processes. These include embryonic development, parturition initiation, and resumption of ovarian activity. In domestic ruminants, the secretion of uterine PGF2 is crucial for the cyclical regression of the corpus luteum. PGF2 metabolizes into PGFM (13,14-dihydro-15-keto-PGF2) during lung passage. PGFM, having a longer half-life in peripheral circulation than PGF2, serves as a useful analytical marker for PGF2 activity.

PGFM’s utility extends to non-invasive pregnancy monitoring in both urine and fecal samples. Fecal PGFM analysis is especially promising for pregnancy diagnosis in various animal species, including captive and free-ranging felids, pandas, and others. The DetectX® PGFM ELISA Kit, with its sensitivity and specificity, is an invaluable tool for researchers and veterinarians studying reproductive physiology and conducting pregnancy diagnostics across a range of species.