Two-Step Co-Immunoprecipitation (TIP) Enables an Efficient and Highly Selective Isolation of Native Protein-Complexes

ABSTRACT: Co-immunoprecipitation (co-IP) is the most frequently used technique for the isolation of protein-protein (PPI) or protein-nucleic acid interactions (PNI) under native expression conditions. A major disadvantage of co-IP is the abundance of non-specific binders that can impede downstream applications. To overcome this limitation, we developed the two-step co-IP (TIP) that significantly improves the purification of native protein-containing complexes. TIP can be applied with a broad range of specific mono- and polyclonal antibodies. Using TIP we purified IKKalpha- and CD95- interacting proteins in primary human T cells, detected all major binders by mass spectrometry analysis and identified two novel CD95-associated proteins.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): T Cell, Cell Culture, Blood

SUBMITTER: Uwe Warnken  

LAB HEAD: Martina Schnoelzer

PROVIDER: PXD006700 | Pride | 2018-01-11

REPOSITORIES: Pride