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Differential Labeling of Cell-surface and Internalized Proteins after Antibody Feeding of Live Cultured Neurons
11:56

Differential Labeling of Cell-surface and Internalized Proteins after Antibody Feeding of Live Cultured Neurons

Published on: February 12, 2014

Immunofluorescence labeling of cell surface antigens in Dictyostelium

Alexandre Vernay1, Pierre Cosson

  • 1Department of Cell Physiology and Metabolism, Geneva Faculty of Medicine, Centre Médical Universitaire, 1 rue Michel Servet, Geneva 4 CH1211, Switzerland. Alexandre.Vernay@unige.ch

BMC Research Notes
|August 14, 2013
PubMed

Insights

Fixation and permeabilization for immunolocalization can cause loss of cell surface antigens. An optimized protocol is proposed to label both surface and intracellular antigens without significant loss.

Area of Science:

  • Cell Biology
  • Immunology
  • Biochemistry

Background:

  • Immunolocalization techniques commonly involve cell fixation and permeabilization before antibody application.
  • These standard procedures can interfere with the detection of certain cellular targets.

Purpose of the Study:

  • To investigate the impact of fixation and permeabilization on cell surface antigen detection.
  • To develop an improved protocol for simultaneous labeling of surface and intracellular antigens.

Main Methods:

  • Assessing a cell surface protein in fixed Dictyostelium cells.
  • Evaluating antigen extraction during standard fixation and permeabilization protocols.
  • Developing and testing an optimized immunolabeling protocol.

Main Results:

  • Standard permeabilization procedures significantly extract abundant cell surface antigens in fixed cells.
  • The degree of antigen loss is procedure- and antigen-dependent.
  • The optimized protocol successfully labels both surface and intracellular antigens.

Conclusions:

  • Standard immunolocalization protocols can lead to artifactual loss of cell surface antigens.
  • An optimized protocol minimizes antigen loss, enabling accurate detection of both surface and intracellular targets.
  • This improved method enhances the reliability of immunolocalization studies.
Abstract

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