Click to add Text Sample Preparation for Mass Spectrometry Sermin Tetik, PhD Marmara University July 2015, New Orleans
Click to add Text An overwiev Introduction Sample Lysis and Protein Extraction Protein Enrichment Protein Quantitation and Staining Peptide Clean Up
Click to add Text Introduction Proper sample preparation for MS-based analysis is critical step in the proteomics workflow.
Click to add Text Introduction Mass spectrometry allows for the analysis of samples ranging in mass from ,000 daltons, in attomole through nanomole quantites.
Click to add Text Introduction The most successful proteomics Labs Sample Preparation InstrumentationSoftware
Click to add Text Proper Sample Preparation Means Better Results Proper Sample Preparation Means Better Results Two significant problems in MS analysis First, the ionization techniques used for large molecules work Second, mass spectrum from a complex mixture is very difficult to fully analyze
Click to add Text Proper Sample Preparation Means Better Results Proper Sample Preparation Means Better Results
Click to add Text Discovery vs. Targeted Proteomic Analysis Discovery vs. Targeted Proteomic Analysis A succsessful proteomics experiment requires integration of the right sample preparation, instrumentation and software.
Click to add Text Discovery vs. Targeted Proteomic Analysis Discovery vs. Targeted Proteomic Analysis Discovery proteomics maximizes protein identification. Targeted proteomics strategies limit the number of features.
Click to add Text Targeted Proteomic Analysis Targeted Proteomic Analysis Targeted proteomics experiments are quantified less than 100 proteins with very high precision, sensitivity, specifity and throughput....are used in Pharmaceutical and Diagnostic applications
Click to add Text Mass Specrometry Workflows Mass Specrometry Workflows
Click to add Text Mass Specrometry Workflow Mass Specrometry Workflow
Click to add Text Sample Lysis and Protein Extraction
Click to add Text All living organisms contain proteolytic enzymes for protein catabolism. Sample Lysis and Protein Extraction
Click to add Text Phosphatases are another class of enzymes that are liberated during cell lysis. 80% occur'ng on serine 20% occur'ng on threonine 1% on threonine residues Sample Lysis and Protein Extraction
Click to add Text Sample Lysis and Protein Extraction
Click to add Text Abundant Protein Depletion
Click to add Text Abundant Protein Depletion
Click to add Text Abundant Protein Depletion
Click to add Text Protein Enrichment Biochemical activity Post-translaional modification (PTM)
Click to add Text Protein Enrichment Shematic summary of a standard immunoprecipitation assay
Click to add Text Protein Enrichment Recent advances in MS technology, in combination with phosphoprotein enrichment using immobilized metal affinity chromatography, have resulted in greater solution of the phosphoproteome.
Click to add Text Active Site Protein Labeling and Enrichment Western blot and MS workflows for targeted capture and analysis of enzymes
Click to add Text Protein Clean Up DialysisDesaltingConcentrating Salts or detergents
Click to add Text Protein Clean Up
Click to add Text Protein Quantitation and Staining In-gel digestion SDS-PAGEextracting
Click to add Text Protein Quantitation and Staining
Click to add Text Protein Quantitation and Staining
Click to add Text Protein Digestion
Click to add Text Protein Digestion
Click to add Text Protein Digestion
Click to add Text Protein Digestion
Click to add Text Protein Digestion
Click to add Text Low-abundant Proteins Post Translationally Modified Peptides 1. Enrichment 2. Clean up Peptide Enrichment
Click to add Text Peptide Enrichment
Click to add Text Peptide Enrichment
Click to add Text Peptide Enrichment
Click to add Text Peptide clean up C18 spin Column C4 column Tips
Click to add Text Peptide clean up
Click to add Text Thank You