ANALYTICAL TECHNIQUES: ELECTROPHORESIS

ELECTROPHORESIS

- Migration of charge particles in an electric field

- Separate proteins on the basis of their electric charge densities

- The process of separating the charged constituents of a sample by means of an electrical current.

Western Blotting

It is a method used to separate, detect and identify one or more proteins in a complex mixture.

Iontophoresis

- Migration of small ions,

- It is produced by electromigration causes the

movement of the charged ions due to low electric potential.

Zone Electrophoresis

- Migration of charged macromolecules in a porous support (paper. Cellulose acetate or agarose gel).

- Used for separating charged particles such as nucleic acids, proteins, and biopolymers.

DENSITOMETER

detecting system

• density, darkening of the degree of light, fraction of incident light transmitted

Support materials

Cellulose acetate

Agarose Gel

Polyacrylamide gel

Cellulose acetate

Cellulose acetylated with acetic anhydride- Acetic anhydride reacts with amines to form amide bond derivatives.

• Separates serum proteins into 5 bands

Agarose Gel

DNA separations

I. Purified fraction of agar

II. 10 -15 bands

Polyacrylamide gel

routinely used for protein analysis

Separates proteins with more movement of buffer and solvent relative to their fixed support.

Electrophoretogram

It is the result of electrophoresis consisting of separated strands of a macromolecule

Polyacrylamide gel

It is routinely used for protein analysis

Capillary electrophoresis

Separation is performed in narrow-bore fuse silica capillaries