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Immunogold Electron Microscopy

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.

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Updated: Feb 25, 2026

Specific Labeling of Mitochondrial Nucleoids for Time-lapse Structured Illumination Microscopy

Specific Labeling of Mitochondrial Nucleoids for Time-lapse Structured Illumination Microscopy

Published on: June 4, 2020

Labeling of Membrane Complexes for Electron Microscopy.

Francesca Gubellini^1,2,3, Rémi Fronzes^4,5,6,7

¹Unite´ G5 Biologie structurale de la sécrétion bactérienne, Institut Pasteur, 25-28 rue du Docteur Roux, 75015, Paris, France. francesca.gubellini@pasteur.fr.

Methods in Molecular Biology (Clifton, N.J.)

|July 30, 2017

Summary

This summary is machine-generated.

Labeling protein subunits within large complexes, particularly membrane proteins, is difficult. This study presents successful labeling strategies for structural characterization of these challenging molecular machines.

Keywords:

Antibody Electron microscopy Gold Labeling Membrane complexes Negative stain Tag

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Mitochondria and Endoplasmic Reticulum Imaging by Correlative Light and Volume Electron Microscopy

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Area of Science:

Biochemistry
Structural Biology
Molecular Biophysics

Background:

Localizing specific subunits within large protein complexes is a significant challenge.
Membrane protein complexes present unique difficulties due to subunit amphipathicity and modularity, complicating genetic manipulation and purification.

Purpose of the Study:

To present effective labeling approaches for the structural characterization of large membrane protein complexes.
To address the challenges associated with studying the architecture of complex biological machinery.

Main Methods:

Development and application of various labeling techniques.
Genetic manipulation strategies for subunit targeting.
Purification protocols for maintaining complex integrity.

Main Results:

Demonstrated successful application of labeling strategies in structural studies.
Overcame difficulties in genetic manipulation and purification of membrane complexes.
Enabled precise localization of subunits within complex structures.

Conclusions:

The presented labeling approaches are effective for the structural characterization of large membrane complexes.
These methods provide solutions for studying the intricate organization of membrane machineries.