LC-MS/MS analysis was used for protein identification of immunoprecipitation. 20 uL eluted samples of immunoprecipitation were separated by 12.5% SDS-PAGE and each gel lane was cut in consecutive gel slices followed by destaining twice using NH4HCO3/ACN. Proteins were digested in-gel with DTT, IAA and trypsin. Digested peptides in the gel slices were then desalted using StageTips with C18 Cartridge (Sigma). Peptides extracts for each gel slice were analyzed by High Performance Liquid Chromatography (HPLC) using EASY column (Fisher Scientific, USA) coupled with a Q-Exactive mass spectrometer (Thermo Finnigan). Resulting MS/MS spectra were searched against the predicted Arabidopsis proteome (35386 entries, 20191018) with maxquant software (version 2.3).
[doi:10.25345/C5GF0N70B]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: FPB1, Shortgun, coIP
Principal Investigators: (in alphabetical order) |
Lin Zhang, Shanghai Normal University, China |
Submitting User: | zhanglin2017 |
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