The Mouse 12-Hydroxyeicosatetraenoic Acid (12-HETE) ELISA Kit is a highly sensitive and specific assay designed for the quantitative detection of 12-HETE in biological samples, including serum, plasma, and cell culture supernatants. 12-HETE, a metabolite of arachidonic acid produced through the 12-lipoxygenase (12-LOX) pathway, plays a critical role in inflammation, cancer, and cardiovascular research.
Key Features
- High Sensitivity and Specificity
- Detects picogram-level concentrations of 12-HETE.
- Exhibits negligible cross-reactivity with related eicosanoids.
- Broad Sample Compatibility
- Suitable for serum, plasma, tissue homogenates, and cell culture supernatants.
- Rapid and Reliable
- Optimized for a straightforward workflow with results in under 4 hours.
- Colorimetric Detection
- Utilizes a highly sensitive chromogenic substrate for accurate quantification.
- Comprehensive Kit Components
- Includes pre-coated ELISA plates, standards, buffers, and detection reagents.
Applications
- Inflammation Research
- Study 12-HETE’s role as a pro-inflammatory mediator in chronic inflammatory diseases (NIH Inflammation Research).
- Cancer Studies
- Investigate the involvement of 12-HETE in tumor growth, angiogenesis, and metastasis (PubMed Cancer Articles).
- Cardiovascular Research
- Analyze 12-HETE’s contribution to atherosclerosis and hypertension (FDA Cardiovascular Research).
- Metabolic and Lipidomics Studies
- Explore the role of 12-HETE in metabolic syndromes and lipid signaling pathways (CDC Lipid Research).
Scientific Background
12-HETE is derived from arachidonic acid through the action of the 12-lipoxygenase enzyme. It acts as a signaling molecule influencing various physiological and pathological processes, including:
- Inflammation: Promotes leukocyte chemotaxis and cytokine release.
- Angiogenesis: Facilitates blood vessel formation in cancer.
- Oxidative Stress: Regulates redox balance in cells.
- Cardiovascular Health: Modulates vascular tone and platelet aggregation.
For more on 12-HETE’s role, explore resources from NCBI or PubMed.
Assay Principle
- Specific Capture:
- The plate is pre-coated with antibodies specific to 12-HETE.
- 12-HETE in the sample competes with a biotinylated 12-HETE standard for binding.
- Signal Amplification:
- Streptavidin-HRP conjugate binds to the biotinylated standard.
- Colorimetric Detection:
- The HRP substrate reacts to produce a measurable color change proportional to the 12-HETE concentration.
- Quantification:
- Absorbance is measured at 450 nm, and 12-HETE levels are calculated using a standard curve.
Kit Components
- Pre-coated 96-well ELISA plate.
- Biotinylated 12-HETE standard.
- Streptavidin-HRP conjugate.
- Wash buffer and assay diluents.
- Substrate solution and stop solution.
- Comprehensive user manual.
Performance Characteristics
| Parameter | Specification |
|---|---|
| Sensitivity | < 10 pg/mL |
| Assay Range | 10 pg/mL – 5000 pg/mL |
| Intra-Assay CV | < 8% |
| Inter-Assay CV | < 10% |
Workflow
- Prepare all reagents, standards, and samples as outlined in the manual.
- Add standards and samples to the wells and incubate.
- Wash the plate and add biotinylated 12-HETE standard.
- Add streptavidin-HRP conjugate and incubate.
- Add substrate solution and measure absorbance at 450 nm.
- Use the standard curve to calculate 12-HETE concentrations.
For detailed protocols, refer to FDA ELISA Guidelines or NIH Assay Procedures.
Research Applications
- Chronic Inflammatory Diseases
- Measure 12-HETE levels in diseases like asthma, rheumatoid arthritis, and inflammatory bowel disease.
- Cancer Biomarker Studies
- Evaluate 12-HETE as a potential biomarker for cancer progression and therapeutic response.
- Drug Development
- Screen and evaluate inhibitors of the 12-LOX pathway.
- Lipidomics Research
- Quantify eicosanoid profiles in metabolic and cardiovascular disorders.
Advantages of the Mouse 12-HETE ELISA Kit
- High Throughput: Process up to 96 samples in a single run.
- Enhanced Accuracy: Strict quality controls ensure reliable results.
- Time-Efficient: Minimal hands-on time with a rapid workflow.
Conclusion
The Mouse 12-HETE ELISA Kit is an indispensable tool for studying the role of 12-HETE in inflammation, cancer, and cardiovascular diseases. Its high sensitivity, specificity, and ease of use make it ideal for both academic and clinical research applications.
For more information or to place an order, visit NIH Research Resources or FDA Diagnostic Tools.