ELECTROPHORESIS

INTRODUCTION
Arne Tiselius
• Father of electrophoresis
• Arne Tiselius (Sweeden,1902-
1971)
• Nobel prize in 1948 for
chemistry “for his research on
electrophoresis and adsorption
analysis ,especially for his
discoveries concerning complex
nature of serum protein”
WHAT IS ELECTROPHORESIS?
• Electro=Electric; phoresis= Migration;Carry accross.
• Migration of charged particles or molecules in a medium
under the influence of an applied electric field
• Usually done with gels formed in tubes, slabs, or on a flat
bed.
WHAT IS ELECTROPHORESIS?
• Charged molecule moves to their counter charge electrode
but electric field removed before it reaches the electrode
• Movement of the charged particle is retarded with the
addition a polymeric gel so that a sufficient time is available
for resolving the sample.
Principle
• In an electrical field charged molecules and particles migrate to the
opposite charge.
• Usually in aqueous solution(Buffer).
• Different molecules and particles in the mixture are migrate at
different speeds: separated into single fractions(bands)
• An electric field is applied to molecules and as they are electrically
charged themselves it results in a force acting upon them.
• greater the charge of the molecule = greater the force applied by the
electrical field
Factors affecting ELECTROPHORESIS

• Magnitude of its charge

Inherent • Charge density
• Molecular weight
Factors • Shape

• Solution pH
External • Electric field
• Solution viscosity
Environment • Temperature
TYPES OF ELECTROPHORESIS
1) Zone Electrophoresis
Paper Electrophoresis
Gel Electrophoresis
Thin Layer Electrophoresis
Cellulose acetate Electrophoresis
2) Moving Boundary Electrophoresis
Capillary Electrophoresis
Isotachophoresis
Isoelectric Focussing
Immuno Electrophoresis
PROCESS
Gel Electrophoresis
• Use of a gelatinous material.
• The gel acts as a support medium
• Used to separate proteins or nucleic acids
GEL TYPES
• Starch-Rarely used •
• Polyacrylamide-Protein, small nucleic acid fragments
• Agarose-Nucleic acids, large proteins
• Cellulose acetate-Proteins
Buffer
• Provides ions in solution for electrical conductivity.
• Prevents the pH changing.
Thus they determine;
• Type of charge on solute
• Extent of ionization of solute
• Electrode towards which the solute will migrate
PROCESS BEHIND GEL ELECTROPHORESIS

Force macromolecules through a porous matrix (gel)

DNA forced to move through matrix by placing gel in

electric field

DNA will flow from negative pole to

Gel acts like a sieve to impede migration

PROCESS BEHIND GEL ELECTROPHORESIS
Key to separating DNA molecules

Larger molecules are strongly impeded by the gel

than smaller molecules

Smaller molecules: faster migration

Shorter fragments farther from origin point

CLINICAL APPLICATION
CONCLUSION
• It involves separation of a components in a sample by the differential
rate of migration of ions by attraction or repulsion in an applied
electric field
• Under the influence of an electric field these charged particles
migrate either to the cathode or to the anode, depending on the
nature of their net charge