Comprehensive Proteome and PTM Analysis
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Corresponding Organization :
Other organizations : Broad Institute
Protocol cited in 15 other protocols
Variable analysis
- Oxidation of methionine
- Protein N-terminal acetylation
- Phosphorylation of serine, threonine and tyrosine residues
- Diglycine modification of lysine residues
- Epsilon-acetylated lysine
- Protein identification
- Protein quantification
- PTM-site localization
- Carbamidomethylation as fixed modification
- Trypsin/P as the digestion enzyme
- Maximum of 3 labeled amino acids and 2 missed cleavages per peptide
- Mass tolerance for precursor ions (20 ppm for first search, 6 ppm for main search)
- Fragment ion mass tolerance (20 ppm)
- Maximum FDR of 1% for protein, peptide and PTM-site identification
- Requirement of 2 or more unique/razor peptides for protein identification
- Requirement of a ratio count of 2 or more for protein quantification per replicate measurement
- Requirement of a localization probability >0.75 in each of the three replicates for PTM-site localization
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