Shotgun proteomic analysis of cerebrospinal fluid using off-gel electrophoresis as the first-dimension separation.
Shotgun proteomic analysis usually employs multidimensional separations with the first dimension most commonly being strong cation exchange (SCX) liquid chromatography (LC). SCX-LC is necessarily a serial process for preparation of multiple samples. Here, we apply a newly available tool, off-gel electrophoresis (OGE), for first-dimension separation of peptide mixtures from digests of cerebrospinal fluid (CSF), a complex and low total protein-containing sample. OGE first-dimension fractionation enabled identification of a total of 156 unique proteins compared to 115 identified in previous work using first-dimension SCX fractionation. OGE can be used to process multiple samples unattended with easy retrieval of the separated fractions. Thus, shotgun analysis using OGE as the first-dimension separation offers a significant advantage both in terms of sample throughput as well as increased numbers of identified proteins.
Waller
Lashanda N
LN
Department of Pharmacology and MUSC Proteomics Center, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
Shores
Kevin
K
Knapp
Daniel R
DR
D002555:Cerebrospinal Fluid / Q000737:chemistry*; D004586:Electrophoresis / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods*
D011506:Proteins
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