Natalia A Petushkova
Institute of Biomedical Chemistry, Russian Academy of Medical Sciences,
119121, Pogodinskaya St., 10, Moscow
Serbia
Research Article
Optimization of the SDS-PAGE gel slicing approach for identification of human liver microsomal proteins via MALDI-TOF mass spectrometry
Author(s): Natalia A Petushkova, Olesya V Larina, Aleksey L Chernobrovkin, Oksana P Trifonova, Yulia S Kisrieva, Andrey V Lisitsa, Irina I Karuzina, Irina I Karuzina and Alexander I ArchakovNatalia A Petushkova, Olesya V Larina, Aleksey L Chernobrovkin, Oksana P Trifonova, Yulia S Kisrieva, Andrey V Lisitsa, Irina I Karuzina, Irina I Karuzina and Alexander I Archakov
Proteomics often exploits sequential cutting of the SDS-PAGE gel lane into slices with subsequent identification of proteins by peptide mass fingerprinting (PMF). In this paper, the influence of slice thickness on protein identification was investigated. Following the separation of the human liver microsomal fraction, the 37 to 75 kDa range of the gel lane was cut into 20 or 40 slices of 0.5 mm and 0.25 mm thickness, respectively. Identification of proteins after trypsinolysis was performed by MALDI-TOF mass spectrometry. A twofold reduction of slice thickness did not influence the number of peaks in PMF-spectra. It was established that at a tolerance of 0.15 Da the number of identified proteins in the 40 slices series exceeded by more than twofold the number of identifications in 20 slices series. It was shown that the decrease of slice thickness leads to a consi.. View More»
DOI: 10.4172/jpb.1000211
