Proteomics Tools
Molecular Biology Tools
Separation & Display Tools
Protein Identification Tools
Protein Structure Tools
Microsequencing
Edman Sequencing
Microsequencing
Generates sequence info from N terminus
Commonly done on low picomolar amounts of protein (5-50 ng)
Newer techniques allow sequencing at the femtomolar level (100 pg)
Up to 20 residues can be read
Allows unambiguous protein ID for 8+ AA
Relatively slow, modestly expensive
Protein ID by MS and 2D gel
Protein ID by MS and 2D gel
Requires gel spots to be cut out (tedious)
Ideal for high throughput (up to 500 samples per day)
Allows modifications to be detected
MS allows protein identification by:
Intact protein molecular weight
Peptide fingerprint molecular weights
Sequencing through MS/MS
Protein ID Protocol
Typical Results
401 spots identified
279 gene products
Confirmed by SAGE, Northern or Southern
Confirmed by amino acid composition
Confirmed by amino acid sequencing
Confirmed by MW & pI
MS Analysis Software
