Identification by Mass Spectrometry

Description / Workflow

Protein identification by high-resolution mass spectrometry in total or fractionated biological extracts. LC-MS analysis enables highly accurate measurement of peptide masses generated by enzymatic digestion (MS), followed by fragmentation to obtain sequence information (MS/MS). The resulting data are then compared against one or more sequence databases using dedicated search engines.

Applications

• Large-scale protein identification
• Identification in complex mixtures
• Sequence coverage analysis
• Identification of interaction partners of a target protein

Requirements / Constraints

• Availability of a sequence database
• Prior purification steps before analysis (to be performed by the requesting team: co-immunoprecipitation, GST pull-down, precipitation using peptides or oligonucleotides immobilized on a solid support, etc.)
• Inclusion of appropriate control conditions
• Monitoring of bait protein enrichment level

Technologies

• PMF (Peptide Mass Fingerprinting)
• LC-MS/MS
• Bottom-up proteomics
• Top-down proteomics

Sites