Samenvatting Natuurkunde Course 3

Natuurkunde Course 3

Gel electrophoresis
Gel electrophoresis is a method for separation and analysis of
-macromolecules (NDA, RNA and proteins) and their fragments
-based on their size and charge under influence of an electric field




 Zone electrophoresis
-paper electrophoresis
 serum proteins
-gel electrophoresis
protein, DNA, RNA
-thin layer electrophoresis
amino acids
-cellulose acetate electrophoresis
clinical diagnostics: haemoglobin, serum proteins for monoclonal gammapathies, urine
proteins, isoenzymes, lipo and glycoproteins.
 Moving boundary electrophoresis
-capillary electrophoresis
forensic sciences: DNA, ink
-isotachophoresis
analytical chemistry: separation/ characterisation ionic compounds
-isoelectric focussing
protein separation on isoelectic point (pl)
-immuno electrophoresis
 protein separation/ characterisation using antibodies

,Electrophoresis
Necessary for electrophoresis
 Electric field, electrodes
 Matrix, polyacrylamide or agarose
 Buffer, electrolyt
 Charged bio-molecules
Charged biomolecules, this depends on:
-pH value
-effective charge
Others ions which are bound can shield the charge of the biomolecule
 Size
Depends on hydrated radius bounded water-molecules dipole-ion bond

, Motion and velocity
 Motion can be uniformly
-velocity v is constant there will be no motion acceleration
 Uniformly accelerated
-motion velocity v is not constant there will be a motion acceleration
*highly charged molecules move faster towards the electrode than molecules with a lesser
charge!
*smaller molecules migrate faster than large molecules!

Force