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Split Green Fluorescent Protein System to Visualize Effectors Delivered from Bacteria During Infection
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Published on: May 24, 2018

Immunolocalization of pathogen effectors.

Eric Kemen1, Kurt Mendgen, Ralf T Voegele

  • 1The Sainsbury Laboratory, John Innes Centre, Norwich, UK.

Methods in Molecular Biology (Clifton, N.J.)
|March 2, 2011
PubMed
Summary
This summary is machine-generated.

Polyclonal antibodies allow protein detection in cells and subcellular structures. This method, immunolocalization, is versatile for studying various pathosystems, even those lacking molecular tools.

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Microscopy

Background:

  • Protein detection is crucial for understanding cellular functions and interactions.
  • Existing molecular tools may not be applicable to all biological systems, particularly complex pathosystems.
  • Immunolocalization offers a powerful alternative for protein detection in diverse biological contexts.

Purpose of the Study:

  • To provide a comprehensive guide on performing immunolocalization for protein detection.
  • To detail methods for obtaining high-quality polyclonal antibodies.
  • To outline sample preparation and protein detection techniques for microscopy.

Main Methods:

  • Generation and validation of high-quality polyclonal antibodies.
  • Detailed protocols for sample preparation for both light and electron microscopy.
  • Application of immunolocalization techniques for cellular and subcellular protein detection.

Main Results:

  • Successful application of immunolocalization across various pathosystems.
  • Demonstration of protein detection at both cellular and subcellular resolutions.
  • Adaptability of the methods for systems not amenable to genetic manipulation.

Conclusions:

  • Immunolocalization using polyclonal antibodies is a robust technique for protein detection.
  • The presented methods are valuable for studying protein localization in diverse biological systems.
  • This approach overcomes limitations of molecular tools in certain experimental setups.