| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Sensitivity |
Assay Principle
Sandwich ELISA
This kit employs a two-site sandwich ELISA to quantitate 8-iso-PGF2α in human samples. The microplate is pre-coated with anti-8-iso-PGF2α capture antibody. Samples and standards bind the capture antibody; biotinylated detection antibody and Streptavidin-HRP are applied sequentially; TMB substrate generates colorimetric signal at 450 nm proportional to 8-iso-PGF2α concentration. Sensitivity: 15 pg/mL; range: 31.25–2,000 pg/mL.
Performance Data
| Assay Type | Sandwich ELISA |
|---|---|
| Species Reactivity | Human (Homo sapiens) |
| Target Analyte | 8-iso-Prostaglandin F2α (8-iso-PGF2a; isoprostane) |
| Sensitivity (LLOD) | 15 pg/mL |
| Detection Range | 31.25–2,000 pg/mL |
| Sample Types | ⚠ Not specified — confirm with manufacturer before use |
| Plate Format | 96-well, available as 48T or 96T |
| Kit Stability | ≤5% activity loss within stated shelf life |
| Manual | Available (PDF, see Documents tab) |
Standard Kit Components
| Component | Description |
|---|---|
| Microplate | 96-well pre-coated with capture antibody (12×8-well strips) |
| Standards | Recombinant 8-iso-PGF2a protein for standard curve |
| Detection Antibody | Biotinylated anti-8-iso-PGF2a detection antibody |
| Streptavidin-HRP | Enzyme conjugate for signal amplification |
| TMB Substrate | 3,3′,5,5′-Tetramethylbenzidine chromogenic substrate |
| Stop Solution | Sulfuric acid stop solution |
| Wash Buffer | Concentrated wash buffer (dilute before use) |
| Sample Diluent | For sample preparation and dilution |
Scientific Background
8-iso-Prostaglandin F2α (8-iso-PGF2α; also 8-epi-PGF2α or isoprostane iPF₂α-III) is produced by non-enzymatic, free radical-mediated peroxidation of arachidonic acid in membrane phospholipids. Unlike enzymatic prostaglandins, isoprostane synthesis is driven directly by reactive oxygen species (ROS) and is independent of COX enzyme activity. 8-iso-PGF2α is widely recognized as one of the most reliable biomarkers of in vivo oxidative stress and lipid peroxidation. Elevated levels are validated in cardiovascular disease, diabetes, Alzheimer's disease, cancer, and acute lung injury. Measurable in plasma, urine, and tissue samples.
Safety & Handling
Research Use Only (RUO). Not for diagnostic or therapeutic applications. TMB substrate and stop solution are corrosive — wear appropriate PPE. Sodium azide may be present — avoid ingestion.
Related Products
Browse additional ELISA kits and immunoassay reagents in the BioHippo catalog.
8-iso-PGF2α is considered a gold-standard oxidative stress biomarker because: (1) it is produced exclusively by non-enzymatic, ROS-driven lipid peroxidation — not by COX enzymes; (2) it is chemically stable and measurable in plasma, urine, and BAL fluid; (3) levels are elevated across diverse oxidative stress-associated diseases; and (4) it is minimally affected by dietary arachidonic acid intake, unlike some other eicosanoids.
Compatible sample types are not explicitly listed by the manufacturer. Typical applications use serum, plasma, urine, or tissue homogenate. Consult the product manual (Documents tab) or contact the manufacturer for confirmed validated matrices. Urine samples may require solid-phase extraction (SPE) prior to ELISA.
Normal human plasma 8-iso-PGF2α levels range approximately 35–200 pg/mL — well within the detection range (31.25–2,000 pg/mL) and far above the 15 pg/mL LLOD. This provides ample signal margin for samples near the lower end of the normal range. Patients with elevated oxidative stress may show levels significantly exceeding 200 pg/mL — prepare 2–5-fold dilutions for such samples.
Prepare an 8-point standard curve by 2-fold serial dilution of the provided 8-iso-PGF2α standard from 2,000 pg/mL to 31.25 pg/mL, plus a zero standard (buffer blank). Run standards and samples in duplicate. Fit using a 4-parameter logistic (4PL) model for accurate concentration interpolation.
Store the complete kit at 2–8°C. The manufacturer guarantees ≤5% activity loss within the stated shelf life under recommended storage conditions. Avoid freezing kit components unless specifically instructed in the manual. Once the plate is opened, return unused strips to the sealed bag with desiccant.
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