Co-IP Mass Spectrometry Analysis

Co-IP samples of sGFP and CjWRKY1-sGFP proteins were obtained as described above. Co-immunoprecipitated samples were separated by SDS-PAGE and silver stained (Supplementary Fig. S2). Protein bands were extracted, and proteins were digested in-gel with MS grade modified trypsin (Promega). The extracted peptides were subjected to LTQ LC/MS/MS (Thermo Fisher Scientific). The collected ion spectrum data were analysed with the help of Mr. Y. Watanabe (Proteomic Facility Lab., Grad. Sch. Biostudies, Kyoto Univ.) using MassMatrix version 2.4.2 (www.massmatrix.net/)38 (link).

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Yamada Y, & Sato F. (2016). Tyrosine phosphorylation and protein degradation control the transcriptional activity of WRKY involved in benzylisoquinoline alkaloid biosynthesis. Scientific Reports, 6, 31988.

Publication 2016

MS grade modified trypsin LTQ LC/MS/MS

Ms ms Peptides Promega Proteins Sds page Silver Trypsin

Corresponding Organization :

Other organizations : Kyoto University

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Variable analysis

independent variables

Co-IP samples of sGFP and CjWRKY1-sGFP proteins

dependent variables

Proteins separated by SDS-PAGE and silver stained
Protein bands extracted and digested with trypsin
Extracted peptides subjected to LTQ LC/MS/MS

control variables

MS grade modified trypsin (Promega) used for in-gel protein digestion

controls

No positive or negative controls were explicitly mentioned in the provided information.

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