title: Combination of acid labile detergent and C18 Empore™ disks for improved identification and sequence coverage of in-gel digested proteins.
abstract: A protocol for improved extraction of peptides from in-gel protein digests, using a combination of the acid labile surfactant, sodium deoxycholate (SDC) and C18 Empore™ membranes, is presented. This approach results in better mass spectrum quality, higher numbers of identified peptide peaks and improved identification scores compared to standard tryptic digestion protocols, or protocols using only SDC or only C18 Empore™ disks. The advantages of the new protocol are demonstrated for two different types of samples: Merino wool intermediate filament proteins and Elaeis guineensis (oil palm) mesocarp proteins.
authors.affiliation: AgResearch Ltd, Private Bag 4749, Christchurch 8140, New Zealand.
authors.forename: Henning
authors.initials: H
authors.lastname: Koehn
authors.forename: Benjamin
authors.initials: B
authors.lastname: Lau
authors.forename: Stefan
authors.initials: S
authors.lastname: Clerens
authors.forename: Jeffrey E
authors.initials: JE
authors.lastname: Plowman
authors.forename: Jolon M
authors.initials: JM
authors.lastname: Dyer
authors.forename: Umi Salamah
authors.initials: US
authors.lastname: Ramli
authors.forename: Santanu
authors.initials: S
authors.lastname: Deb-Choudhury
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D028023:Arecaceae / Q000737:chemistry; D003840:Deoxycholic Acid / Q000737:chemistry; D003902:Detergents / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods*; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000295:instrumentation / Q000379:methods*; D010940:Plant Proteins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014935:Wool / Q000737:chemistry*
chemical_list: D003902:Detergents; D010940:Plant Proteins; D003840:Deoxycholic Acid