Advantages and Disadvantages
Provides a hard-copy record of separation
Allows facile quantitation
Separation of up to 9000 different proteins
Highly reproducible
Gives info on Mw, pI and post-trans modifications
Inexpensive
Limited pI range (4-8)
Proteins >150 kD not seen in 2D gels
Difficult to see membrane proteins (>30% of all proteins)
Only detects high abundance proteins (top 30% typically)
Time consuming
2D Gel Software
Capillary Electrophoresis
Capillary Electrophoresis
Capillary Zone Electrophoresis (CZE)
Separates on basis of m/z ratio
Capillary Gel Electrophoresis (CGE)
Separates by MW and m/z ratio
Capillary Isoelectric Focusing (CIEF)
Separates on basis of pI
2-Dimensional Electrophoresis (2D-CE)
Separates using tandem CE methods
Chromatography
Size Exclusion (size)
Reverse Phase (hphob)
Ion Exchange (charge)
Normal Phase (TLC)
Affinity (ligand)
HIC (hydrophobicity)
2D Chromatography
Ciphergen Protein Chips
Ciphergen Protein Chips
Hydrophobic (C8) Arrays
Hydrophilic (SiO2) Arrays
Anion exchange Arrays
Cation exchange Arrays
Immobilized Metal Affinity (NTA-nitroloacetic acid) Arrays
Epoxy Surface (amine and thiol binding) Arrays
Ciphergen Protein Chips
Protein Arrays
Different Kinds of Protein Arrays
Detection by: SELDI MS, fluorescence, SPR, electrochemical, radioactivity, microcantelever
