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Enrichment of NICD4 interacting proteins by three different methods.

ABSTRACT: There are several methods to bring down the bait and interactors from the cell lysates, each with its own advantages and disadvantages. Recently, the most commonly used method is one step AP, TAP and proximity-labeling. In order to analyze the Strengths and weaknesses of the three methods, fusion proteins were constructed by NICD4 with FLAG, SFB (S tag-2x Flag tag-SBP tag) and TurboID respectively, NICD4 and its interactors were obtained by purification. By analyzing these proteins, we believe that SFB-TAP is the most reliable and effective method to identify interacting proteins.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Acute Leukemia

SUBMITTER: bian weixiang

LAB HEAD: Xu Li

PROVIDER: PXD031772 | Pride | 2022-08-12

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	CSFB_NICD4_1.mzid.gz	Mzid
	CSFB_NICD4_1.mzid_CSFB_NICD4_1.MGF	Mzid
	CSFB_NICD4_1.raw	Raw
	CSFB_NICD4_1.sf3	Other
	CSFB_NICD4_2.mzid.gz	Mzid

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Publications

Protocol for establishing a protein-protein interaction network using tandem affinity purification followed by mass spectrometry in mammalian cells.

Bian Weixiang W Jiang Hua H Feng Shan S Chen Junjie J Wang Wenqi W Li Xu X

STAR protocols 20220719 3

Identification of protein interactors is fundamental to understanding their functions. Here, we describe a modified protocol for tandem affinity purification coupled with mass spectrometry (TAP/MS), which includes two-step purification. We detail the S-, 2×FLAG-, and Streptavidin-Binding Peptide (SBP)- tandem tags (SFB-tag) system for protein purification. This protocol can be used to identify protein interactors and establish a high-confidence protein-protein interaction network based on comput ...[more]

PMID: 35874475

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