Identification of
Protein Spots in Silver-stained Two-dimensional Gels by MALDI-TOF Mass Peptide
Map Analysis
CHEN Ping, XIE Jing-Yun, LIANG Song-Ping*
( College of Life Science, Hunan Normal University, Changsha 410081,
China )
Abstract Protein
spots in silver-stained two dimensional gels were analyzed and identified by
employing an improved procedure of mass spectrometric peptide mapping,
including： i) In-gel reduction, alkylation and
enzymatic digestion； ii) Extraction and desalting by using a
pipette tip containing a small C18 micro-column (ZipTipTM)； iii) Direct MALDI-TOF mass
analysis and protein database searching. The results demonstrate that single
silver-stained protein spots in a 2-DE-gel could be identified rapidly by this
procedure and the use of the ZipTipTM pipette tip could increase
evidently the sensitivity of the MALDI-TOF analysis. By using this the
procedure, 10 protein spots in a silver-stained gel of 2-DE of crude venom of
the spider S.Huwena were analyzed and identified.
Key words MALDI-TOF-MS； proteome；peptide mass fingerprint；2D-PAGE
*Corresponding author： Tel, 86-731-8872556； Fax, 86-731-8861304； e-mail, [email protected]