Methods in molecular biology (Clifton, N.J.), 2021, Vol.2262, p.47
2021
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Details
- Autor(en) / Beteiligte
- Titel
- Precise Characterization of KRAS4B Proteoforms by Combining Immunoprecipitation with Top-Down Mass Spectrometry
- Ist Teil von
- Methods in molecular biology (Clifton, N.J.), 2021, Vol.2262, p.47
- Ort / Verlag
- United States
- Erscheinungsjahr
- 2021
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- eISSN: 1940-6029DOI: 10.1007/978-1-0716-1190-6_3Titel-ID: cdi_pubmed_primary_33977470
- Format
- –
- Schlagworte
- Chromatography, Liquid - methods, Humans, Immunoprecipitation - methods, Neoplasms - metabolism, Neoplasms - pathology, Protein Isoforms, Protein Processing, Post-Translational, Proteome - analysis, Proto-Oncogene Proteins p21(ras) - analysis, Proto-Oncogene Proteins p21(ras) - metabolism, Tandem Mass Spectrometry - methods, Tumor Cells, Cultured