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Imaging proteins in live mammalian cells with biotin ligase and monovalent streptavidin.

Abstract:

This protocol describes a simple and efficient way to label specific cell surface proteins with biophysical probes on mammalian cells. Cell surface proteins tagged with a 15-amino acid peptide are biotinylated by Escherichia coli biotin ligase (BirA), whereas endogenous proteins are not modified. The biotin group then allows sensitive and stable binding by streptavidin conjugates. This protocol describes the optimal use of BirA and streptavidin for site-specific labeling and also how to produ...

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Publication status:
Published

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Publisher copy:
10.1038/nprot.2008.20

Authors


Howarth, M More by this author
Journal:
Nature protocols
Volume:
3
Issue:
3
Pages:
534-545
Publication date:
2008
DOI:
EISSN:
1750-2799
ISSN:
1754-2189
URN:
uuid:2b81cb41-77d1-4004-8844-f94d044c49d2
Source identifiers:
101220
Local pid:
pubs:101220

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