Electrophoresis

ELECTROPHORESIS

migration of the charged particle through a solution under the influence of an external electrical field

ZONE ELECTROPHORESIS

it involves the migration of the charged particle on the supporting media

PAPER ELECTROPHORESIS

It is economical

Easy to use

PAPER ELECTROPHORESIS

Disadvantage: Certain compounds such as protein, hydrophilic molecules cannot be resolved due to the adsorptive and ionogenic properties of paper which results in tailing and distortion of components band

GEL ELECTROPHORESIS

Separation is brought about through molecular sieving technique, based on the molecular size of the substances.

porous gel

acts as a sieve by retarding or, in some cases, by completely obstructing the movement of macromolecules while allowing smaller molecules to migrate freely.

agar

mixture of polysaccharides extracted from seaweeds.

Agarose

chemically basic disaccharide repeating units of 3,6-anhydro-L-galactose

agar and agarose gel

Advantage: It absorbs proteins relatively less when compared to other mediums.

agar and agarose gel

Resolution is less compared to polyacrylamide gels

POLYACRYLAMIDE GEL (PAGE)

It is prepared by polymerizing acrylamide monomers in the presence of methylene-bis-acrylamide to cross link the monomers.

POLYACRYLAMIDE GEL (PAGE)

It is thermostable, transparent, strong and relatively chemically inert. Gels are uncharged and are prepared in a variety of pore sizes.

Native - PAGE

Type of polyacrylamide gel that Separation is based upon charge, size, and shape of macromolecules. It is the original mode of electrophoreses

DENATURED - PAGE

SDS- PAGE

Type of polyacrylamide gel that Separation is based upon the molecular weight of proteins.

GEL ELECTROPHORESIS using STARCH

A suspension of granular starch should be boiled in a buffer to give a clear colloidal suspension.

petroleum jelly

What was used to avoid swelling and shrinking in gel electrophoresis using starch

GEL ELECTROPHORESIS using STARCH

Disadvantage: Variation in pore size from batch to batch

THIN LAYER ELECTROPHORESIS

Studies can be carried out in a thin layer of silica, alumina

THIN LAYER ELECTROPHORESIS

Widely used in combined electrophoretic- chromatography studies in two- dimensional study of proteins and nucleic acid hydrolysates

CELLULOSE - ACETATE ELECTROPHORESIS

It contains 2-3 acetyl groups per glucose unit and its adsorption capacity is less than that of paper

CELLULOSE - ACETATE ELECTROPHORESIS

No tailing of proteins or hydrophilic materials.

CELLULOSE - ACETATE ELECTROPHORESIS

Widely used in analysis of clinical and biological protein samples (albumin and globulins).

zone electrophoresis.

For testing purity of thyroid hormones

Paper chromato-electrophoresis

used to separate free Insulin from plasma proteins.

Electrophoresis

is also used for separation of carbohydrates and vitamins.