"Spot transfer", elution and comigration with known proteins allows accurate transferral of protein identifications between distinct two-dimensional electrophoretic systems.
We report a simple method, designated "spot transfer", where known proteins are excised and eluted from a two-dimensional (2-D) gel run on one gel system to transfer identification of the proteins to a different 2-D gel system by comigration with a comparable sample. In one experiment, 8 of 16 proteins eluted from an isoelectric focusing (IEF) 2-D gel, of the format described for the Celis human keratinocyte database (Celis, J. E. et al., Electrophoresis 1993, 14, 1091-1198), were found to comigrate with proteins in a human T lymphoma (JURKAT) whole cell lysate run using the Millipore Investigator 2-D system. The method should have general utility in allowing the exchange of protein identifications between investigators and could be used to standardize gel loci used in 2-D gel protein databases.
Dean
D P
DP
Department of Molecular Genetics and Protein Chemistry, Pfizer, Inc., Groton, CT 06340.
Cronan
M T
MT
Merenda
J M
JM
Gardner
J P
JP
Connelly
P A
PA
Celis
J E
JE
D001345:Autoradiography / Q000379:methods; D002460:Cell Line; D002478:Cells, Cultured; D016208:Databases, Factual; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D016399:Lymphoma, T-Cell; D008715:Methionine / Q000378:metabolism; D009363:Neoplasm Proteins / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D013462:Sulfur Radioisotopes; D014407:Tumor Cells, Cultured
D009363:Neoplasm Proteins; D011506:Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine