DIGE compatible labelling of surface proteins on vital cells in vitro and in vivo.
Efficient methods for profiling of the cell surface proteome are desirable to get a deeper insight in basic biological processes, to localise proteins and to uncover proteins differentially expressed in diseases. Here we present a strategy to target cell surface exposed proteins via fluorescence labelling using CyDye DIGE fluors. This method has been applied to human cell lines in vitro as well as to a complex biological system in vivo. It allows detection of fluorophore-tagged cell surface proteins and visualisation of the accessible proteome within a single 2-D gel, simplifying subsequent UV MALDI-MS analysis.
Mayrhofer
Corina
C
Department of Clinical Pathology, Medical University of Vienna, Vienna, Austria.
Krieger
Sigurd
S
Allmaier
Günter
G
Kerjaschki
Dontscho
D
D000818:Animals; D001859:Bone Neoplasms / Q000378:metabolism; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D012516:Osteosarcoma / Q000378:metabolism; D020543:Proteome; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D005456:Fluorescent Dyes; D008565:Membrane Proteins; D020543:Proteome