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Methods Mol Biol ; 2564: 317-323, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36107351

RESUMO

Protein-protein interactions play a vital role in the cellular physiology of living organisms. Among several available approaches, co-immunoprecipitation (co-IP) has emerged as a reliable method to investigate such interactions. The underlying principle is to retrieve a bait protein from a protein extract using bait-specific antibodies and thereby indirectly capture the interacting partner proteins. However, bait-specific antibodies are not always available, and the genetic fusion of a peptide tag offers an alternative. An extensive range of peptide tags and the tag-specific antibodies are available nowadays. Fluorescent proteins are widely used protein tags for co-IP experiments. In this chapter, we describe a method to co-immunoprecipitate the fluorescently tagged candidate protein with its interacting partners from the crude plant cell extracts using green fluorescent protein (GFP)-trap magnetic beads.


Assuntos
Ligante de CD40 , Células Vegetais , Anticorpos , Extratos Celulares , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Imunoprecipitação , Peptídeos , Células Vegetais/metabolismo
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