NDLI: Bimolecular affinity purification (BAP): tandem affinity purification using two protein baits.

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Bimolecular affinity purification (BAP): tandem affinity purification using two protein baits.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Maine, Gabriel N. Gluck, Nathan Zaidi, Iram W. Burstein, Ezra
Description	Country affiliation: United States Author Affiliation: Maine GN ( Department of Clinical Pathology, William Beaumont Hospital, Royal Oak, MI 48073, USA.)
Abstract	The tandem affinity purification (TAP) procedure was pioneered in yeast for the purpose of purifying and characterizing protein complexes. While affinity purification is relatively easy to perform, nonspecific protein interactions can plague the identification of true interacting partners of the given bait utilized in the purification. To alleviate this problem, two sequential affinity purification steps are employed in the TAP procedure. Since its inception in yeast, TAP has gone through many adaptations and has been employed multiple times in diverse organisms, including mammalian systems. In all these approaches, two out of many possible affinity moieties are employed and are usually expressed as a fusion polypeptide in the amino or carboxyl-terminal region of the protein bait. In this protocol, we describe a variation on the TAP procedure in which the affinity moieties are placed on two different proteins of a molecular complex to isolate or detect components present in the complex. This variation, which we refer to as bimolecular affinity purification (BAP), is suited for the identification of specific molecular complexes marked by the presence of two known components.
File Format	HTM / HTML
ISSN	19403402
e-ISSN	15596095
DOI	10.1101/pdb.prot5318
Journal	Cold Spring Harbor Protocols
Issue Number	11
Volume Number	2009
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2009-11-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Clinical Laboratory Techniques Chromatography, Affinity Proteins Metabolism Cell Line Cloning, Molecular Isolation & Purification
Content Type	Text
Resource Type	Article
Subject	Biochemistry, Genetics and Molecular Biology

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Bimolecular Affinity Purification (BAP): Tandem Affinity Purification Using Two Protein Baits

IgG affinity capture of TAP-tagged protein complexes from cell extracts: affinity purification step 1.

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Affinity capture of TAP-tagged protein complexes: affinity purification step 2.

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SAINT: probabilistic scoring of affinity purification-mass spectrometry data.

Affinity purification of protein complexes.

Growing and harvesting TAP-tagged yeast cells.

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Bimolecular affinity purification (BAP): tandem affinity purification using two protein baits.

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Bimolecular Affinity Purification (BAP): Tandem Affinity Purification Using Two Protein Baits

IgG affinity capture of TAP-tagged protein complexes from cell extracts: affinity purification step 1.

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Bimolecular affinity purification (BAP): tandem affinity purification using two protein baits.