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This document describes the Routine 3.1 PVDF sequencer methods used with the Hewlett-Packard G1005A N-terminal Protein Sequencing System. These methods implement Edman degradation chemistry for analyzing proteins electroblotted to PVDF membranes in the nanomole to low picomole range.
The manual provides detailed step-by-step instructions and volume specifications for three specific methods: Routine 3.1 PVDF (Column), Routine 3.1 PVDF (Converter), and Routine 3.1/3.5 PTH Std. It also includes a Column Prep method to wash the sequencer column prior to sample loading.
| Applicable Sample Range | nanomole to low picomole range |
|---|---|
| Routine 3.1 PVDF Column Method Time | 45.4 min (2722.1 sec) |
| Routine 3.1 PVDF Cycle 1 Time | 61.2 min (3670.1 sec) |
| Routine 3.1 PVDF Converter Method Time | 33.1 min (1988.1 sec) |
| Routine PTH Std Delivery Volume | 10 pmol/100 |
| Converter Step 2 R4 Delivery Volume | approximately 70 pl (+5 pl) |
It provides an initial sample cleanup and performs a double coupling on the column.
Copy Steps 7, 8, and 9 to the clipboard, run from the clipboard, remove the conversion flask after Step 9, unscrew it, and measure the delivered volume.