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Advanced Insights into Nuclear Factor NF-kappa-B p65 Subunit (NFKB3) ELISA Kits: Applications and Methodologies

Nuclear factor NF-kappa-B (NF-κB) is a transcription factor critical in regulating immune responses, inflammation, and cell survival. The p65 subunit, also known as NFKB3, plays a pivotal role in the canonical NF-κB signaling pathway. Its activation involves translocation to the nucleus, where it modulates the expression of target genes involved in immunity, inflammation, and cellular stress responses (NIH NF-kappa-B Research).

Importance of Measuring NF-kappa-B p65

Quantifying NF-kappa-B p65 levels is essential in understanding its involvement in:

What is an NF-kappa-B p65 ELISA Kit?

An NF-kappa-B p65 (NFKB3) ELISA kit is a laboratory tool designed to detect and quantify NF-kappa-B p65 in various biological samples, such as cell lysates, plasma, and serum. These kits provide a specific and sensitive method for assessing NF-kappa-B p65 activity and expression levels in research and clinical studies.

Features and Benefits of NF-kappa-B p65 ELISA Kits

  1. High Sensitivity: Detects low concentrations of NF-kappa-B p65, suitable for diverse applications.
  2. Specificity: Utilizes antibodies highly specific to the p65 subunit, ensuring accurate results.
  3. Ease of Use: Includes pre-coated microplates and optimized reagents for streamlined workflows (FDA Diagnostic Standards).
  4. Versatility: Compatible with a range of biological samples and experimental designs.
  5. Reproducibility: Delivers consistent results with high inter- and intra-assay precision (NIH Rigor and Reproducibility Guidelines).

Applications of NF-kappa-B p65 ELISA Kits

  1. Inflammatory Research:
    • Investigating the role of NF-kappa-B p65 in inflammatory pathways.
    • Evaluating therapeutic agents targeting NF-kappa-B signaling (NIH Inflammation Research).
  2. Cancer Studies:
    • Assessing NF-kappa-B activity in tumor microenvironments.
    • Monitoring the efficacy of anti-cancer drugs targeting NF-kappa-B signaling pathways (PubMed Cancer Resources).
  3. Immunology:
    • Studying NF-kappa-B p65’s role in immune cell activation and differentiation.
    • Identifying biomarkers for autoimmune and immunodeficiency disorders (CDC Immunology Research).
  4. Cellular Stress Research:
    • Examining NF-kappa-B p65 activation under oxidative and endoplasmic reticulum stress.
    • Investigating its role in apoptosis and cell survival mechanisms (NIH Cellular Stress Studies).

Protocol Overview for NF-kappa-B p65 ELISA Kits

  1. Sample Preparation: Process cell lysates, plasma, or serum samples according to the kit’s instructions to ensure protein stability.
  2. Reagent Preparation: Dilute standards, controls, and reagents as specified in the kit manual.
  3. Assay Procedure:
    • Add samples and standards to the microplate wells.
    • Incubate with specific antibodies and enzyme conjugates.
    • Add substrate solution and measure colorimetric changes for quantification.
  4. Data Analysis: Generate a standard curve and calculate NF-kappa-B p65 concentrations using appropriate software (NIH ELISA Methodology).

Selecting the Right NF-kappa-B p65 ELISA Kit

  • Validation: Choose a kit validated for your sample type and research application.
  • Sensitivity and Range: Select a kit with detection limits appropriate for your experimental needs.
  • Specificity: Verify that the antibodies used are specific to the NF-kappa-B p65 subunit.
  • Reproducibility: Opt for kits with proven consistency in inter- and intra-assay variability (FDA Quality Standards).

Future Directions in NF-kappa-B Research

  1. Therapeutic Development: Targeting NF-kappa-B p65 in the treatment of cancer, autoimmune diseases, and chronic inflammation.
  2. Biomarker Discovery: Establishing NF-kappa-B p65 as a diagnostic and prognostic biomarker in various diseases.
  3. Systems Biology: Leveraging NF-kappa-B signaling studies for understanding complex cellular networks and disease mechanisms (NIH Systems Biology Resources).

Conclusion

The NF-kappa-B p65 subunit (NFKB3) is a critical regulator of immune responses, inflammation, and cell survival. ELISA kits for NF-kappa-B p65 provide powerful tools for quantifying this protein, enabling advancements in understanding its role in health and disease. For further information, explore resources from trusted organizations such as the National Institutes of Health, Centers for Disease Control and Prevention, and FDA.

 

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