title: RP-HPLC prefractionation and its application in expressional proteomics analysis of an in vitro viral infection model.
abstract: Prefractionation of complex protein mixtures is an efficient method for increasing the separation power of 2-DE. RP-HPLC has been successfully utilized as a prefractionation method prior to 2-DE. Here we describe the optimization of an efficient RP-HPLC method for prefractionation of baby hamster kidney cell solubilized proteins. A step gradient elution of acetonitrile was optimized and collected fractions were further examined by SDS-PAGE and 2-DE. By utilizing this method an effective increase in separation power of 2-DE is accomplished. Moreover, we describe the application of this method to expressional proteome analysis of a virally infected cell model.
authors.affiliation: Protein Chemistry Unit, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran. Behrouz-vaziri@pasteur.ac.ir
authors.forename: Behrouz
authors.initials: B
authors.lastname: Vaziri
authors.forename: Mehdi
authors.initials: M
authors.lastname: Rahimpour
authors.forename: Naser
authors.initials: N
authors.lastname: Eslami
authors.forename: Ahmad
authors.initials: A
authors.lastname: Fayaz
authors.forename: Heshmat
authors.initials: H
authors.lastname: Rahimian
mesh_terms: D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006224:Cricetinae; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D066298:In Vitro Techniques; D008954:Models, Biological; D020543:Proteome / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D011818:Rabies / Q000378:metabolism; D011820:Rabies virus; D012967:Sodium Dodecyl Sulfate; D014764:Viral Proteins / Q000302:isolation & purification*
chemical_list: D020543:Proteome; D014764:Viral Proteins; D012967:Sodium Dodecyl Sulfate