PUBLICATION

Correlative light and electron microscopic detection of GFP-labeled proteins using modular APEX

Authors
Ariotti, N., Hall, T.E., Parton, R.G.
ID
ZDB-PUB-170523-13
Date
2017
Source
Methods in cell biology   140: 105-121 (Chapter)
Registered Authors
Hall, Thomas, Parton, Robert G.
Keywords
APEX, CLEM, Electron microscopy, GFP-binding peptide, Green fluorescent protein, Light microscopy
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Ascorbate Peroxidases/metabolism*
  • Cell Line, Tumor
  • Green Fluorescent Proteins/metabolism*
  • Microscopy, Electron/methods*
  • Subcellular Fractions/metabolism
  • Zebrafish
PubMed
28528629 Full text @ Meth. Cell. Biol.
Abstract
The use of green fluorescent protein (GFP) and related proteins has revolutionized light microscopy. Here we describe a rapid and simple method to localize GFP-tagged proteins in cells and in tissues by electron microscopy (EM) using a modular approach involving a small GFP-binding peptide (GBP) fused to the ascorbate peroxidase-derived APEX2 tag. We provide a method for visualizing GFP-tagged proteins by light and EM in cultured cells and in the zebrafish using modular APEX-GBP. Furthermore, we describe in detail the benefits of this technique over many of the currently available correlative light and electron microscopy approaches and demonstrate APEX-GBP is readily applicable to modern three-dimensional techniques.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping