Mass Spectrometry enables determination of the relative molecular masses of small molecules as well as of large biomolecules such as proteins and peptides. The ABSCIEX Triple TOF 5600 system housed at RCB the is most sensitive, high-resolution LC-MS/MS mass spectrometer with high mass accuracy and equipped with smart speed acquisition. The Triple TOF 5600 has both ESI and APPI ionization sources and is is hyphenated with a Shimadzu UFLC for LC-MS analysis of small molecules and also with an Eksigent Nano LC-Ultra system for use in demanding proteomic applications.
2-D Gel Electrophoresis
2D gel electrophoresis is one of the most widely used tools in proteomic applications. This technique allows separating the proteins in first dimension according to the charge (Isoelectric focusing, IFE) followed by separating the focused proteins in the second dimension according to the molecular weight by SDS-PAGE. The proteins can be visualized by coomassie Blue, silver stain and fluorescent dyes. The two-dimensional array produces spots corresponding to single proteins in the sample. Using the this techniques thousands of protein can be separated and molecular properties like pI, molecular weight and protein amount can be determined. This spot can be further characterized using MALDI-TOF-TOF Mass spectrometry. Fluorescent-based labeling and image quantification can accurately estimate cellular regulation of proteins. We have Bio-Rad 2D Gel-electrophoresis with Molecular ChemiDoc XRS imaging system.
Proteomic Facility in ATPC
The Advanced Technology Platform Centre (ATPC) of Bioscience cluster is armed with AB SCIEX TOF/TOF 5800 System with ekspert nanoLC 400 and EKSpot MALDI Spotter for comprehensive proteomics applications both qualitative and quantitative. This system facilitates the identification and analysis of the total complement of proteins and other biomolecules. The system also capable of performing to the identification of intact protein, protein/peptide identification and characterization, protein biomarker research, PTM identification and analysis, De novo sequencing, MALDI imaging, gel based and non-gel based applications, quantitative proteomics including labelled (iTRAQ, SILAC etc.), lipid and polymer analysis.
Protein Sequencer (PPSQ-33A)
Central instrumentation facility (CIF) of RCB is equipped with an automated peptide/protein sequencing system, PPSQ-33A from Shimadzu Corporation. The PPSQ-33A consists of an Edman reaction unit, an online PTH-amino acid Analyzer with LC pump and UV detector, and Windows95 based software.