Vinay, Protein sequence analysis consists of the following steps
1.Digest the protein to the peptides. Remember, mass spectrometry gets limited sequence data from complete proteins. However, they can analyse peptides.
2. Distinguish peptides on reverse phase column using acetonitrile gradient. Use column 75 µm in diameter and use acetic acid in the solvents.
3.Put ionised peptides in vapor phase through passing the column eluate, that consists of peptides and solvent. Use chromatography to include molecules into a mass spectrometer.
4.Calculate mass of peptides.
5.Fragment peptides.
6.Measure the mass of fragments from peptides.
7.Use fragment mass data to find out the sequence of the peptide.
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