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6 years ago in Data Analysis By Vinay Kumar
Protein analysis using mass spectroscopy
How do I analyse protein using mass spectroscopy? Also I have heard that it is not possible to completely sequence data for proteins. Why so?
All Answers (3 Answers In All)
By Sapna C K Answered 6 years ago
1. Proteins does not digest
2. Peptides too small pass through reverse phase column
3. Peptides too large still to gel and do not elute from the column/tube
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By Krishan Pancholi Answered 6 years ago
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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By Divya Answered 6 years ago
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