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Immunohistochemical Analysis of Expression, Phosphorylation, and Nuclear Translocation of NF-κB Proteins in Human

Davide Vecchiotti1, Daniela Verzella1, Daria Capece1

  • 1Department of Biotechnological and Applied Clinical Sciences (DISCAB), University of L'Aquila, L'Aquila, Italy.

Methods in Molecular Biology (Clifton, N.J.)
|July 8, 2021
PubMed

Insights

This study details immunohistochemistry (IHC) for identifying NF-κB proteins in human tissues. The methods cover both formalin-fixed paraffin-embedded and frozen tissue types for accurate antigen localization.

Area of Science:

  • Biomedical Science
  • Cell Biology
  • Histology

Background:

  • Immunohistochemistry (IHC) is a crucial technique for visualizing antigen localization in tissues.
  • Understanding cellular and subcellular antigen distribution is vital in biological research.
  • NF-κB proteins play significant roles in cellular responses and disease.

Purpose of the Study:

  • To provide a comprehensive guide for identifying and localizing NF-κB proteins in human tissues.
  • To detail the application of immunohistochemistry for NF-κB protein detection.
  • To cover protocols for both formalin-fixed paraffin-embedded and frozen tissue samples.

Main Methods:

  • Utilizing immunohistochemistry (IHC) staining techniques.
  • Employing specific targeting reagents labeled with reporter molecules.
  • Applying methods to formalin-fixed paraffin-embedded and frozen human tissue sections.

Main Results:

  • Successful identification of NF-κB protein presence and localization within human tissue samples.
  • Demonstration of IHC's efficacy in visualizing cellular and subcellular antigen distribution.
  • Detailed protocols for reproducible results in tissue analysis.

Conclusions:

  • Immunohistochemistry is an effective method for localizing NF-κB proteins in human tissues.
  • The described techniques are applicable to both FFPE and frozen tissue sections.
  • This provides a valuable resource for researchers studying NF-κB in human pathology and biology.

Keywords:
AntibodiesAntigen retrievalDetectionFixationIdentification (localization)MicroscopeNF-κB

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