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CURREN
T
LIMITATI
ONS
CROP SYSTEMS
TISSUE AND
CELL
ISOLATIONS
PROTEIN
EXTRACTION
PROTEIN SEPARATION, DIGESTION &
QUANTITATION
DATABASE
SEARCH &
MAPPING TO
GENOME
DATA
VALIDATION
AND
INTERPRETAT
ION
metabolite profiling
High throughput
enzymatic assay
Bioinformatics tools
and databases
BIOMARKERS
DEVELOPMENT
CURRENT
LIMITATIONS
FUTURE DIRECTIONS
- Diversity of systems
Model crop species for
proteomics studies
- Selection of cell types
- Laser microdissection
- Fluorescent-activated
cell sorting (FACS)
- Highly abundant
proteins
- Detergent compatible
with 2-D gel & labelling
- Depletion by solvent,
antibody & fractionation
- Sequential extraction
- CPLLs
- Costs of DIGE and labelling approaches
- Detection & quantitation of low abundant proteins
- Characterization of protein modifications
- Label-free proteomics
- Peptide enrichment
- Selected reaction monitoring (SRM)
- High accuracy and sensitivity mass spectrometers
- Poor annotation
- High sequence
redundancy
- Curated databases
- Identification and
quantitation by unique
peptide
- Multihit and
normalization strategies
- Availability of
bioinformatics
tools
- Access to validation
tools
- High-throughput
enzymatic
assays
- Costs for large scale
proteomics
- Label-free proteomics
- Orthoproteomics
- Large scale
comparative
proteomics
- Targeted approach
(SRM)
in contrasting genotypes
New Microsoft PowerPoint Presentation.pptx

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New Microsoft PowerPoint Presentation.pptx

  • 1. CURREN T LIMITATI ONS CROP SYSTEMS TISSUE AND CELL ISOLATIONS PROTEIN EXTRACTION PROTEIN SEPARATION, DIGESTION & QUANTITATION DATABASE SEARCH & MAPPING TO GENOME DATA VALIDATION AND INTERPRETAT ION metabolite profiling High throughput enzymatic assay Bioinformatics tools and databases BIOMARKERS DEVELOPMENT CURRENT LIMITATIONS FUTURE DIRECTIONS - Diversity of systems Model crop species for proteomics studies - Selection of cell types - Laser microdissection - Fluorescent-activated cell sorting (FACS) - Highly abundant proteins - Detergent compatible with 2-D gel & labelling - Depletion by solvent, antibody & fractionation - Sequential extraction - CPLLs - Costs of DIGE and labelling approaches - Detection & quantitation of low abundant proteins - Characterization of protein modifications - Label-free proteomics - Peptide enrichment - Selected reaction monitoring (SRM) - High accuracy and sensitivity mass spectrometers - Poor annotation - High sequence redundancy - Curated databases - Identification and quantitation by unique peptide - Multihit and normalization strategies - Availability of bioinformatics tools - Access to validation tools - High-throughput enzymatic assays - Costs for large scale proteomics - Label-free proteomics - Orthoproteomics - Large scale comparative proteomics - Targeted approach (SRM) in contrasting genotypes