Edman Experimental Procedure

Liquid samples for Protein Sequence Analysis are spotted onto Glass Fiber Filters that are pretreated with Biobrene. Biobrene is used to ensure that the sample is retained within the sequenator.

Samples that have been electroblotted to PVDF membranes are submitted directly to analysis without further processing, although, samples that are contaminated by glycine from the electroblotting procedure may be washed with water prior to the analysis. Excess glycine will interfere with the analysis for the first several cycles of analysis.

All sequence analyses include a separate chromatogram of known PTH amino acid standards for calibration purposes. These standards are separated to greater than 98% resolution by reverse phase HPLC. Retention times of the standards are used for peak identification and quantification.  Customized reports are generated using the SequencePro software giving chromatograms of each cycle, repetitive yield graphs and histograms. A blank HPLC trace is included in all experiments to evaluate background signal. Blank runs and standard runs are included with the raw data in your final report.

The sequencing procedure is automated on the Procise 492 and controlled by the Procise Sequencer software (version 5.1). All analyses are visually inspected and confirmed by PCL staff trained in sequence analysis prior to reporting.

You will receive a final report in pdf format by email. Unused samples cannot be returned unless by special arrangement.