| Field | Specification |
|---|---|
| Mfr No | |
| Alternative Names | 8-epi-PGF2a |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
| Storage |
Features & Benefits
Human 8-epi-prostaglandin F2alpha (8-epi-PGF2α) ELISA Kit has high sensitivity and excellent specificity for detection of Human 8-epi-PGF2a. No significant cross-reactivity or interference between Human 8-epi-PGF2a and analogues was observed.
Background
The isoprostanes are a family of eicosanoids of non-enzymatic origin produced by the random oxidation of tissue phospholipids by oxygen radicals. Isoprostanes appear as artifacts in tissue and plasma samples which have undergone oxidative degradation during prolonged or improper storage. They also appear in the plasma and urine under normal conditions and are elevated by oxidative stress. At least one of the isoprostanes, 8-isoprostane (8-epi PGF2α), has been shown to have biological activity. It is a potent pulmonary and renal vasoconstrictor1 and has been implicated as a causative mediator of hepatorenal syndrome and pulmonary oxygen toxicity.2 8-Isoprostane has been proposed as a marker of antioxidant deficiency and oxidative stress and elevated levels have been found in heavy smokers.3 8-Isoprostane levels are also a relative indicator of sample integrity for lipid-containing samples such as serum, plasma, and whole cell preparations.
This 8-epi-prostaglandin F2alpha (8-epi-PGF2α) ELISA kit is validated for use with Cell culture supernatants, Serum, Plasma, Other biological fluids. Samples should be collected, processed, and stored correctly to preserve analyte integrity — avoid repeated freeze-thaw cycles and centrifuge to remove particulates before use. Dilute samples exceeding the kit's detection range using the supplied assay diluent. Hemolytic, icteric, or lipemic samples may affect assay performance and should be tested with caution.
This is a sandwich ELISA kit employing an HRP (horseradish peroxidase)-conjugated secondary antibody paired with a TMB (3,3′,5,5′-tetramethylbenzidine) colorimetric substrate. In the sandwich format, the target analyte 8-epi-prostaglandin F2alpha (8-epi-PGF2α) captured on the microplate surface is detected by the conjugated antibody, generating a colorimetric signal proportional to analyte concentration. The reaction is stopped and absorbance measured at 450 nm on a standard microplate reader.
The complete protocol, from sample addition to final plate reading, requires approximately 3–5 hours. This includes two incubation periods (analyte binding and detection antibody steps), intermediate wash cycles to remove unbound material, 15–30 minutes of TMB substrate development, and final stop-solution addition before absorbance reading. Exact timing will vary with experience level and the number of samples processed in parallel.
Required equipment: (1) a microplate spectrophotometer capable of reading absorbance at 450 nm (reference wavelength 540–570 nm recommended for background correction); (2) precision single-channel or multichannel pipettes; (3) a plate washer or multichannel aspirator; (4) a microcentrifuge for sample clarification; and (5) a 37°C incubator or stable room-temperature environment. No fluorescence or luminescence reader is required — standard colorimetric plate readers are fully compatible with this kit.
This ELISA kit is formally validated for Human. Cross-reactivity with species not listed in the specification has not been independently characterized. Variability in protein sequence homology across species means that performance in unlisted species cannot be guaranteed without additional validation. For cross-species detection requirements or non-standard sample matrices, please contact BioHippo support or refer to the manufacturer's technical team for guidance.
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