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Studying changes in global protein expression in a cell or tissue system

Which analyses do I require?. Studying changes in global protein expression in a cell or tissue system (Classical proteomics ‘fishing’). Larger amounts of protein available (>50 ug) ( note/ if any identification of proteins using mass spec will be required then ~ 300 ug

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Studying changes in global protein expression in a cell or tissue system

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  1. Which analyses do I require? Studying changes in global protein expression in a cell or tissue system (Classical proteomics ‘fishing’) • Larger amounts of protein available (>50 ug) • (note/ if any identification of proteins using • mass spec will be required then ~ 300 ug • preparative gels will be required) • b) Molecular weight range > 20 kDa • Limited amount of protein available (< 25 ug) • Molecular weight range <30 kDa • Hydrophobic / membrane-bound proteins • Unlimited amounts of protein available > 0.5 mg) • Molecular weight range required 1 kDa (peptides)- 300 kDa ProteinChip Array Technology (SELDI TOF MS) AND 2D-PAGE / DIGE ProteinChip Array Technology (SELDI TOF MS) 2D-PAGE / DIGE

  2. Which analyses do I require? Protein: Protein interactions or Antibody-Capture Experiments (affinity chromatography) • Limited amount of protein available • Molecular weight range < 100 kDa • Large numbers or High-throughput screening • Target protein identity is not required • Larger amounts of protein available (> 0.5 mg) • b) Identification of the target protein is required Protein or antibody immobilised on IDM affinity beads (Ciphergen) ProteinChip Array Technology (SELDI TOF MS) RS100 arrays 1D or 2D-LC Electrospray-QTOF Mass Spectrometry Note/ These procedures are a specialised analyses and you will need to contact us to discuss the possibility of any collaborations

  3. Which analyses do I require? Identification, Sequencing and Characterisation of proteins (including proteins excised from polyacrylamide gels) Post-translational modifications of proteins (PTM) Identification of proteins excised from PAGE gels Tryptic digestion and peptide analysis by MALDI TOF MS The identification of sites of PTM For e.g. phosphorylation, glycosylation, nitration, nitrosylation etc. Glycan analysis Mass Mapping Identification Tryptic digestion and peptide analysis by ESI-Q-TOF MS Mass Mapping identification fails (insufficient peptide masses / keratin contamination / not in database) Release of Glycans (PNGase F or Hydrazinolysis) Optional analyses Protein sequencing using electrospray quadrupole time of flight mass spectrometry (ESI-QTOF MS) MALDI and / or ESI quadrupole time of flight mass spectrometry (MALDI / ESI -QTOF MS)

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