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Methods Mol Biol. 2017;1575:175-187. doi: 10.1007/978-1-4939-6857-2_10.

Antibody Validation by Immunoprecipitation Followed by Mass Spectrometry Analysis.

Author information

1
Science for Life Laboratory, Drug Discovery and Development Platform & School of Biotechnology, KTH-Royal Institute of Technology, Tomtebodavägen 23A, 17165, Solna, Sweden. helena.persson@scilifelab.se.
2
Structural Genomics Consortium, Department of Biochemistry and Biophysics, Karolinska Institutet, Solna, Sweden.
3
Terrence Donnelly Center for Cellular & Biomolecular Research, University of Toronto, Toronto, Canada.
4
Centre for Molecular Medicine, Rheumatology Unit, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, Solna, Sweden.

Abstract

We describe a mass spectrometry-based approach for validation of antibody specificity. This method allows validation of antibodies or antibody fragments, against their endogenous targets. It can assess if the antibody is able to bind to its native antigen in cell lysates among thousands of other proteins, DNA, RNA, and other cellular components. In addition, it identifies other proteins the antibody is able to immunoprecipitate allowing for the assessment of antibody specificity and selectivity. This method is easily scalable, adaptable to different cell lines and conditions and has been shown to be reproducible between multiple laboratories.

KEYWORDS:

Antibody fragments; Antibody validation; Immunoprecipitation; Mass spectrometry

PMID:
28255880
DOI:
10.1007/978-1-4939-6857-2_10
[Indexed for MEDLINE]

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