Proteomics. 2005 Jun;5(9):2417-21.

Two-stage affinity purification for inducibly phosphorylated membrane proteins.

Peirce MJ, Begum S, Saklatvala J, Cope AP, Wait R.

Source

Kennedy Institute of Rheumatology Division, Imperial College, London, UK. m.peirce@imperial.ac.uk

Abstract

Characterisation of tyrosine phosphorylations induced in immune cells in response to inflammatory stimuli may help elucidate the molecular bases of the diversity of immune responses. We have used anti-phosphotyrosine antibodies in combination with cell surface biotinylation in a two-step affinity purification procedure to recover pervanadate-induced tyrosine phosphorylated proteins from sub-cellular compartments, including the cell surface, of murine T cells and macrophages prior to separation by solution-phase isoelectric focussing and one-dimensional gel electrophoresis and identification by tandem mass spectrometry.

PMID:: 15887181; [PubMed - indexed for MEDLINE]

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Two-stage affinity purification for inducibly phosphorylated membrane proteins.

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