- The movement of charged particles in an electrical field resulting in their migration towards the positively charged electrodes is known as electrophoresis.
- Molecules with a net negative charge move towards positively charged electrode, while those with net positive charge move towards negatively charged electrode.
- The rate of migration of ions in an electric field depends on several factors like:-
- Molecular weight, shape, size and net charge of particles.
- Solubility of ions
- Viscosity of the solution
- Magnitude of the current applied.
- Classification based on mode of operation and separation:-
- Moving Boundary
- Zone
- Paper
- Cellulose Acetate
- Gel
- Immunoelectrophoresis
- Electrophoresis Apparatus:-
- Anode and cathode separated by a wall
- Each side filled with a buffer up to same level
- A bridge to connect the anode and cathode
- Example for Serum proteins:-
- Buffer used is barbital(pH: 8.6)
- Being negatively charged they migrate towards anode
- Albumin with maximum charge moves the most distance.
- Gamma globulins with minimum charge move the least distance.
- Clinical applications:-
- Separation of serum proteins, isoenzymes, hemoglobulin, etc.
- Used in western blotting to detect HIV antibodies
- Used in southern blotting to identify DNA, RNA.
- Determination of Immunoglobulin types.