| Literature DB >> 33977470 |
Lauren M Adams1, Caroline J DeHart2, Neil L Kelleher3,4,5.
Abstract
The characterization of biologically relevant post-translational modifications (PTMs) on KRAS4B has historically been carried out through methodologies such as immunoblotting with PTM-specific antibodies or peptide-based proteomic methods. While these methods have the potential to identify a given PTM on KRAS4B, they are incapable of characterizing or distinguishing the different molecular forms or proteoforms of KRAS4B from those of related RAS isoforms. We present a method that combines immunoprecipitation of KRAS4B with top-down mass spectrometry (IP-TDMS), thus enabling the precise characterization of intact KRAS4B proteoforms. We provide detailed protocols for the IP, LC-MS/MS, and data analysis comprising a successful IP-TDMS assay in the contexts of cancer cell lines and tissue samples.Entities:
Keywords: Immunoprecipitation; Intact protein; Isoform; Proteoform; RAS; Top-down mass spectrometry
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Year: 2021 PMID： 33977470 PMCID： PMC8543976 DOI： 10.1007/978-1-0716-1190-6_3
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745