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Protein Identification Using 20-Minute Edman Cycles and Sequence Mixture Analysis

1999; Elsevier BV; Volume: 267; Issue: 1 Linguagem: Inglês

10.1006/abio.1998.2981

1096-0309

William J. Henzel, Jake Tropea, David Dupont,

Microfluidic and Capillary Electrophoresis Applications

We have developed a 20-min Edman cycle and a multiple sample horizontal flow reactor for the sequence analysis of PVDF-electroblotted proteins. The 20-min cycle uses a 12-min C18 phenylthiohydantoin separation. This cycle and separation is compatible with most Applied Biosystems sequencers. Using this rapid cycle, 10 residues on six different proteins can be completed within a 24-h period. We also demonstrate the use of an algorithm that can sort mixture sequences derived from PVDF bands that contain coeluting proteins.