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Protein purification methods

Protein purification methods


Daily thousands of laboratories or research facilities require methods or techniques for the detection and quantitation of proteins. Analytical requirements can range from standard protein detection and characterization methods to clinical diagnostics and testing even drug dosing techniques. However, multiple factors need to be considered when selecting the best approach or method for this purpouse. The following is a list of methods used for the purification and characterization of proteins from various sources. 

General methods to Purify Proteins

  • Protein Isolation using chromatography methods:

o   Ion exchange,

o   Size-exclusion chromatography or gelfiltration,

o   Affinity chromatography,

o   Liquid chromatography with perfusion columns

  • Protein Extraction and Solubilization
  • Protein Concentration Determination Methods
  • Concentrating Protein Solutions
  • Gel electrophoresis
    • Gel Electrophoresis using denaturing conditions
    • Gel Electrophoresis using non-denaturing conditions
    • 2D Gel Electrophoresis
  • Electrofocusing

Methods to analysis Protein Structures

  • X-ray crystallography
  • Protein NMR

Protein-Protein interactions

  • Yeast two-hybrid system
  • Protein-fragment complementation assay
  • Co-immunoprecipitation
  • Affinity purification and mass spectrometry

Protein-DNA interactions

  • ChIP-on-chip
  • Chip-Sequencing
  • DamID (adenine methyltransferase identification)
  • Microscale Thermophoresis

Computational methods

  • Molecular dynamics
  • Protein structure prediction
  • Protein sequence alignment (sequence comparison, incl. BLAST)
  • Protein structural alignment
  • Protein ontology, see gene ontology

Other methods

  • Hydrogen-deuterium exchange
  • Mass spectrometry
  • Protein sequencing
  • Protein synthesis
  • Proteomics
  • Peptide mass fingerprinting
  • Ligand binding assay
  • Eastern blotting
  • metabolic labeling
    • heavy isotope labeling
    • radioactive isotope labeling