Hye Lin Park Author Answered Jul 24, 2022
Department of Botany and Plant Pathology and the Center of Plant Biology, Purdue University, West Lafayette
Thank you for your question. We transiently expressed the YFP-ACS5 in tobacco leaves, and we did the immunoprecipitation to get rid of other tobacco proteins. Without IP, you can not check YFP-ACS5 is phosphorylated by MBP-SINAT2 or not, and we used UB-antibody to confirm the ubiquitination of YFP-ACS5. GFP-antibody is already used for IP process, so we can not use GFP-antibody for check the ubiquitination.