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  • A simple and fast protocol for the protein complex immunoprecipitation (Co-IP) of effector: host protein complexes.

A simple and fast protocol for the protein complex immunoprecipitation (Co-IP) of effector: host protein complexes.

Methods in molecular biology (Clifton, N.J.) (2014-03-20)
Jens Steinbrenner, Matthew Eldridge, Daniel F A Tomé, Jim L Beynon
ABSTRACT

Plant pathogens are responsible for enormous damage in natural and cultured ecosystems. One strategy most pathogenic organisms follow is the secretion of effector proteins that manipulate the host immune system to suppress defense responses. There is considerable interest in finding host targets of pathogen effectors as this helps to shape our understanding of how those proteins work in planta. The presented protocol describes a protein complex immunoprecipitation method aimed at verifying protein-protein interactions derived from protein complementation assays like Yeast-two-Hybrid.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
KOD Hot Start Master Mix, Ready-to-use 2X mixture, containing KOD Hot Start DNA Polymerase, two monoclonal antibodies, ultrapure deoxynucleotides, and reaction buffer with MgSO4, optimized for convenient high fidelity PCR., ready-to-use solution, suitable for PCR
Roche
Anti-GFP, from mouse IgG1κ (clones 7.1 and 13.1)
Roche
cOmplete ULTRA Tablets, Mini, EDTA-free, EASYpack Protease Inhibitor Cocktail, Tablets supplied in foil blister packs.