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    Large-scale immunoprecipitation from fission yeast cell extracts.

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    Authors
    Grallert, Agnes
    Hagan, Iain M
    Affiliation
    CRUK Cell Division Group, Cancer Research UK Manchester Institute, University of Manchester, Manchester M20 4BX, United Kingdom
    Issue Date
    2017-02-01
    
    Metadata
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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.
    Citation
    Large-scale immunoprecipitation from fission yeast cell extracts. 2017, Cold Spring Harb Protoc
    Journal
    Cold Spring Harbor Protocols
    URI
    http://hdl.handle.net/10541/620185
    DOI
    10.1101/pdb.prot091595
    PubMed ID
    28148853
    Type
    Article
    Language
    en
    ISSN
    1559-6095
    ae974a485f413a2113503eed53cd6c53
    10.1101/pdb.prot091595
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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