The N-to-C 4.0 and C-to-N 4.0 sequencer methods enable switching between the N-terminal and C-terminal sequencing chem- istry methods on the Hewlett- Packard column-based protein sequencer. The N-to-C 4.0 method is run when switching from the N-terminal to the C-terminal sequencing chem- istry. The column method pauses the column, while the flask method prepares a coordinated injection with the on-line HPLC to place the proper HPLC solvents in the HPLC binary pumps for the C-terminal chromatography. H N-to-C 4.0 and C-to-N 4.0 Protein Sequencer Methods Technical Note The C-to-N 4.0 method is run when switching from the C-terminal to the N-terminal sequencing chem- istry. The column method washes the column, while the flask method, in addition to washing the flask, prepares a coordinated injection with the on-line HPLC to place the proper HPLC solvents in the HPLC binary pumps for the N-terminal chromatography. These methods are generally run with a Zitex strip in an empty RP/SAX biphasic col- umn, but any column configuration is applicable. No column prepara- tion is required with this method. The methods are: • N-to-C 4.0 (column) • N-to-C 4.0 (flask) • Column wash 4.0 • Flask wash 4.0 The sequence program controls all of the column and flask methods, and well as any cycle exception methods. The N-to-C 4.0 and C-to-N 4.0 sequencer methods do not use a exception method. The Routine 4.0 sequencer methods require the bottle configuration shown. The reagents and solvents are
This document provides technical details on the N-to-C 4.0 and C-to-N 4.0 sequencer methods used with the Hewlett-Packard 241 Protein Sequencer (N+C). These methods enable users to switch between N-terminal and C-terminal sequencing chemistry, utilizing either a column method or a flask method depending on the specific transition required.
The N-to-C 4.0 method is used when switching from the N-terminal to the C-terminal chemistry, and the C-to-N 4.0 method is used when switching from the C-terminal to the N-terminal chemistry.
The sequencer column configuration consists of an empty reverse-phase (RP) sample column mated with an empty strong anion exchange (SAX) column.
No, no column preparation is required when using the N-to-C 4.0 or C-to-N 4.0 sequencer methods.