Detection of epitope-tagged recombinant LRR-RLK proteins in Arabidopsis with Western hybridization. Full-length cDNAs of SERK1, SERK2, SERK3, SERK4, SERK5 and BRI1 were recombined into pB35GWF and pK35GWG. FLAG-tagged fusion proteins, prepared by immunoprecipitation of total membrane protein, and total proteins from leaves of transgenic plants expressing GFP-tagged fusion proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Proteins were transferred to nitrocellulose membrane and epitope-tagged proteins were detected using either (A) anti-FLAG or (B) anti-GFP antibody (diluted 1:3,000) followed by anti-mouse immunoglobulin G (IgG)-horseradish peroxidase (HRP) secondary antibody (diluted 1:10,000).
Gou et al. BMC Genomics 2010 11:19 doi:10.1186/1471-2164-11-19