The OASIS® HLB μElution plate as a one-step platform for manual high-throughput in-gel digestion of proteins and peptide desalting.
Separation or prefractionation of proteins by gel electrophoresis, followed by in-gel proteolytic digest and analysis by MS is a typical standard application in proteomics. For many laboratories, it is not cost-effective to use a robot with all its drawbacks. On the other hand, manual digest is time consuming and not free of error if many samples are processed at the same time. The OASIS® HLB μElution plate allows the handling of 96 samples for protein in-gel digest and peptide desalting in a semiautomatic way. Using multichannel pipettes, solutions are added quickly, and with the use of the positive pressure-96 stand all solutions are removed simultaneously. In this article, a complete and detailed protocol for the use of the OASIS® HLB μElution plate is introduced and its effectiveness is shown with 2D- and 1D-gel samples.
Max-Planck-Institute for Biology of Ageing, Bio-MS Facility, Cologne, Germany. franz@age.mpg.de
Thomas
T
Franz
Xinping
X
Li
D000818:Animals; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D057166:High-Throughput Screening Assays / Q000295:instrumentation* / Q000379:methods*; D010455:Peptides / Q000378:metabolism*; D059748:Proteolysis; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D012965:Sodium Chloride / Q000302:isolation & purification
D029721:Drosophila Proteins; D010455:Peptides; D012965:Sodium Chloride