Journal
CHEMBIOCHEM
Volume 18, Issue 15, Pages 1492-1495Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201700177
Keywords
epitope tags; fluorescence microscopy; fluorescent probes; SpyTag-SpyCatcher; super-resolution microscopy
Funding
- National Institutes of Health [DP2OD008479]
- NIH [R21 EB022798]
- American Heart Association
- National Science Foundation
Ask authors/readers for more resources
Labeling proteins with high specificity and efficiency is a fundamental prerequisite for microscopic visualization of subcellular protein structures and interactions. Although the comparatively small size of epitope tags makes them less perturbative to fusion proteins, they require the use of large antibodies that often limit probe accessibility and effective resolution. Here we use the covalent SpyTag-SpyCatcher system as an epitope-like tag for fluorescent labeling of intracellular proteins in fixed cells for both conventional and super-resolution microscopy. We also applied this method to endogenous proteins by gene editing, demonstrating its high labeling efficiency and capability for isoform-specific labeling.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available