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Large-scale immunoprecipitation from fission yeast cell extracts.

Grallert, Agnes
; 
Hagan, Iain M
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Abstract
We outline immunoprecipitation (IP) procedures to isolate the large quantities of a molecule of interest that are required to identify posttranslational modifications (PTMs) in subsequent targeted mass spectrometry analysis. In situ denaturation by trichloroacetic acid precipitation inhibits the activities of modifying enzymes that could alter the PTM profile to preserve the PTMs on a target of interest throughout the precipitation step. In contrast, isolation of the same molecule with the nondenaturing variation on this IP procedure can maintain associations with partner molecules whose PTMs can also be mapped, albeit with the caveat that modifications could have occurred during the extended IP period.
Authors
Grallert, Agnes
Hagan, Iain M
Affiliation
CRUK Cell Division Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester M20 4BX, United Kingdom
Description
Date
2017-02-01
Publisher
Collections
All Paterson Institute for Cancer Research
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Article
Citation
Large-scale immunoprecipitation from fission yeast cell extracts. 2017, Cold Spring Harb Protoc
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Volume Title
URI
http://hdl.handle.net/10541/620185
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