Automation of micro-preparation and enzymatic cleavage of gel electrophoretically separated proteins.
To achieve high throughput, protein microcharacterization sample preparation must be automated. We describe a cartesian robot capable of processing 32 protein samples in parallel. The system is based on specially designed flow-through reactors for contamination-free reagent delivery and removal. Washing of excised gel pieces, reduction and alkylation, proteolytic cleavage and peptide extraction are performed in these reactors. Compatibility of the system with HPLC peptide separation and Edman degradation as well as with laser desorption mass spectrometry of the unseparated mixture is demonstrated. This is the first report describing automated preparation and processing of multiple protein samples.
Houthaeve
T
T
Protein and Peptide Group, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Gausepohl
H
H
Mann
M
M
Ashman
K
K
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism; D015003:Yeasts / Q000737:chemistry
D010455:Peptides; D011506:Proteins; D014357:Trypsin