Saltar al contenido
Merck
  • Protein and antibody purification followed by immunoprecipitation of MYB and GATA zinc finger-type maize proteins with magnetic beads.

Protein and antibody purification followed by immunoprecipitation of MYB and GATA zinc finger-type maize proteins with magnetic beads.

STAR protocols (2022-06-14)
Isabel Cristina Vélez-Bermúdez, Jorge Enrique Salazar-Henao, Marta Riera, David Caparros-Ruiz, Wolfgang Schmidt
RESUMEN

Co-immunoprecipitation (Co-IP) is a widely used and powerful approach for studying protein-protein interactions in vivo. Here, we describe a protocol for antibody purification and immobilization followed by immunoprecipitation from plant tissue extracts using magnetic beads. The protocol has been used to detect regulators in the Zea mays phenylpropanoid pathway. The protocol is amenable to a variety of downstream assays, including western blotting and mass spectrometry. For complete details on the use and execution of this protocol, please refer to Vélez-Bermúdez et al. (2015).

MATERIALES
Product Number
Marca
Descripción del producto

Sigma-Aldrich
Glicina, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
Seroalbúmina bovina, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
DL-Ditiotreitol solution, BioUltra, Molecular Biology, ~1 M in H2O
Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
Rosetta(DE3) Competent Cells - Novagen, Rosetta host strains are BL21 derivatives designed to enhance the expression of eukaryotic proteins that contain codons rarely used in E. coli.