Multiparameter screen optimizes immunoprecipitation.

Autor:	Xie S; European Research Institute for the Biology of Ageing, University Medical Centre Groningen, Groningen, 9713AV, The Netherlands., Saba L; European Research Institute for the Biology of Ageing, University Medical Centre Groningen, Groningen, 9713AV, The Netherlands., Jiang H; Laboratory of Cellular & Structural Biology, The Rockefeller University, New York, NY 10065, USA., Bringas OR; European Research Institute for the Biology of Ageing, University Medical Centre Groningen, Groningen, 9713AV, The Netherlands., Oghbaie M; European Research Institute for the Biology of Ageing, University Medical Centre Groningen, Groningen, 9713AV, The Netherlands.; Laboratory of Cellular & Structural Biology, The Rockefeller University, New York, NY 10065, USA., Stefano LD; European Research Institute for the Biology of Ageing, University Medical Centre Groningen, Groningen, 9713AV, The Netherlands., Sherman V; High Energy Physics Instrument Shop, The Rockefeller University, New York, NY 10065, USA., LaCava J; European Research Institute for the Biology of Ageing, University Medical Centre Groningen, Groningen, 9713AV, The Netherlands.; Laboratory of Cellular & Structural Biology, The Rockefeller University, New York, NY 10065, USA.
Jazyk:	angličtina
Zdroj:	BioTechniques [Biotechniques] 2024 Apr; Vol. 76 (4), pp. 145-152. Date of Electronic Publication: 2024 Feb 29.
DOI:	10.2144/btn-2023-0051
Abstrakt:	Immunoprecipitation (IP) coupled with mass spectrometry effectively maps protein-protein interactions when genome-wide, affinity-tagged cell collections are used. Such studies have recorded significant portions of the compositions of physiological protein complexes, providing draft 'interactomes'; yet many constituents of protein complexes still remain uncharted. This gap exists partly because high-throughput approaches cannot optimize each IP. A key challenge for IP optimization is stabilizing in vivo interactions during the transfer from cells to test tubes; failure to do so leads to the loss of genuine interactions during the IP and subsequent failure to detect. Our high-content screening method explores the relationship between in vitro chemical conditions and IP outcomes, enabling rapid empirical optimization of conditions for capturing target macromolecular assemblies.
Databáze:	MEDLINE
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