Internal Protein Sequencing:

The Protein Chemistry Laboratory currently operates an Applied Biosystems 173/A Microblotter:

With this technology, the Protein Chemistry Laboratory has the capability of generating internal sequence information from proteins that have blocked N-termini. The protein can be digested with enzymes either in solution, on PVDF, or in an SDS-PAGE gel. The resulting peptides can be are extracted and then separated with the Microblotter's capillary HPLC. The resulting peptide fractions can then be collected utilizing the Microblotter's dynamic solenoid blotting the fractions on a strip of PVDF. After collecting the peptides on the PVDF strip, the peptides can be sequenced using the PROCISE 494.

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• SDS-PAGE and Electroblotting procedure for protein sequencing samples.

• In-Gel Digest procedure for obtaining internal peptide sequences from proteins with blocked N-termini.

• On-PVDF membrane digest procedure for obtaining internal peptide sequences from proteins with blocked N-termini.