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Specifications
- Assay Type Competitive ELISA
- Sample Types Serum, Plasma, Fecal Extracts, Urine, Tissue Culture Media
- Sensitivity 20.8 pg/mL
- Species PGFM is identical across species
- Assay Duration 1.5 Hours
- Samples/Plate 39 in Duplicate
- Readout Colorimetric, 450 nm
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Standard Curve
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Description
Assay Principle:
The PGFM ELISA Kit quantitatively measures PGFM in serum, plasma, fecal extracts, urine, and tissue culture media. The PGFM 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 PGFM 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-rabbit IgG antibody. Add the PGFM peroxidase conjugate and the PGFM polyclonal rabbit antibody. Then incubate the mixture covered at room temperature, shaking for 1 hour. The immunological reaction occurs between the anti-PGFM antibody, the PGFM antigen in the sample or standard, and the PGFM-peroxidase conjugate. As the PGFM concentration in the sample increases, the bound PGFM-peroxidase conjugate decreases, causing a decrease in signal and vice versa.
After the 1-hour incubation, wash away the excess PGFM-peroxidase conjugate and add the TMB substrate. The TMB substrate reacts with the bound PGFM-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the PGFM concentration in the samples.
Background:
Uterine and placental Prostaglandin F2 (PGF2) regulate reproductive and pregnancy-related processes such as embryonic development, parturition initiation, and ovarian activity resumption. In domestic ruminants, uterine tissue is a primary source of PGF2, and secretion of uterine PGF2 is a crucial regulator for the cyclical regression of the corpus luteum. Metabolism of Prostaglandin F2 to PGFM (13,14-dihydro-15-keto-PGF2) occurs during lung passage. PGFM has a longer half-life in peripheral circulation than PGF2 and is a useful analytical marker of PGF2. PGFM is a useful non-invasive marker of pregnancy when measured in both urine and fecal samples. It is a precise, practical method for this application in these matrices. Fecal PGFM analyses may allow pregnancy diagnosis in captive and free-ranging felids, pandas, and other species.