The document describes the N-terminal PVDF 4.0 sequencer methods of HP 241 protein sequencer (N+C)
This document provides technical notes on the N-terminal PVDF 4.0 sequencer methods used with the Hewlett-Packard column-based protein sequencer. The methods implement Edman degradation chemistry for analyzing most protein and peptide samples in the low nanomole to low picomole range loaded onto a Hewlett-Packard PVDF column.
The guide details specific protocols, including 'N-Terminal PVDF 4.0' (total time: 45.7 min), 'N-Terminal PVDF 4.0 (Cycle 1)' (total time: 76.2 min), and 'N-Terminal Flask 4.0' (total time: 34.6 min). It specifies the required bottle configurations, reagent delivery volumes, drying times, and temperature settings for each step of the sequencing process.
| Sample Range | low nanomole to low picomole range |
|---|---|
| Total Time (Column Method) | 45.7 min |
| Total Time (Cycle 1 Method) | 76.2 min |
| Total Time (Flask Method) | 34.6 min |
| Standard Solution Volume | 10 pmol/100 ul |
| Liquid Waste Volume | less than lmL/cycle |
Reagents 1R, 2R, 38R, and the TH-Std are specific for C-terminal sequencing and are not utilized for the N-terminal PVDF 4.0 sequencer methods.
For protein samples, the PVDF column consists of an empty SAX column on top and a RP column on the bottom.
For peptide samples, the PVDF column consists of an empty RP column on top and a SAX column on the bottom.
The Column Prep 4.0 method is used to wash the sequencer column prior to sample loading, which is particularly important for low-level sample quantities.