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Methods Mol Biol. 2017;1657:347-359. doi: 10.1007/978-1-4939-7240-1_27.

Identification of c-di-AMP-Binding Proteins Using Magnetic Beads.

Author information

1
Department of General Microbiology, Georg-August-University Göttingen, Grisebachstr. 8, 37077, Göttingen, Germany.
2
Department of General Microbiology, Georg-August-University Göttingen, Grisebachstr. 8, 37077, Göttingen, Germany. jstuelk@gwdg.de.

Abstract

To identify cytosolic proteins that bind to cyclic di-AMP, a biotinylated analog of the nucleotide is used for protein pull-down experiments. In this approach, biotinylated c-di-AMP is coupled to Streptactin-covered beads. After protein separation using standard SDS-PAGE, the protein(s) of interest are identified by mass spectrometric analyses.

KEYWORDS:

Bacteria; Biotinylated c-di-AMP; Magnetic beads; Protein pull-down

PMID:
28889307
DOI:
10.1007/978-1-4939-7240-1_27
[Indexed for MEDLINE]

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