Abstract
AbstractThis guideline describes parameters and conditions involved in phosphopeptide sample preparation. It covers from the description and preparation of the cells and tissues to the fractionation and specific enrichment of phosphopeptides for MS analysis. The guideline is prepared in order to easily cope with many of the experimental designs used in phosphoproteomic studies. The document is subdivided as follows:1. General features2. Sample processing3. Protein Purification/Fractionation4. Peptide Purification/Fractionation5. Phosphopeptide enrichment
Highlights
In the study of the phosphoproteome, sample preparation for peptide or protein enrichment is an especially critical step. This is achieved using a combination of procedures including HPLC separation of peptides or proteins, gel electrophoresis and enrichment methods such as immunoaffinity or IMAC and TiO2-based purification procedures
This guideline describes parameters and conditions involved in phosphopeptide identification by mass spectrometry
It covers from the description and preparation of the cells and tissues to the fractionation and specific enrichment of phosphopeptides for MS analysis
Summary
In the study of the phosphoproteome, sample preparation for peptide or protein enrichment is an especially critical step. In most cases, this is achieved using a combination of procedures including HPLC separation of peptides or proteins, gel electrophoresis and enrichment methods such as immunoaffinity or IMAC and TiO2-based purification procedures. This guideline describes parameters and conditions involved in phosphopeptide identification by mass spectrometry. It covers from the description and preparation of the cells and tissues to the fractionation and specific enrichment of phosphopeptides for MS analysis. If procedures are repeated more than once, the corresponding section should be repeated (e.g. More than one purification method for phosphopeptide enrichment). This proposal includes some definitions from HUPO-PSI Gel Electrophoresis vs. 1.4, January 2008 and Column Chromatography vs. 1.0, June 2008 documents
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