What is polyacrylamide gel electrophoresis?

Polymerized acrylamide (polyacrylamide) forms a mesh-like matrix suitable for the separation of proteins of typical size. The strength of the gel allows easy handling. Polyacrylamide gel electrophoresis of SDS-treated proteins allows researchers to separate proteins based on their length in an easy, inexpensive, and relatively accurate manner.

How do you polymerize acrylamide?

Pour acrylamide solution for a stacking gel; insert a comb and allow the acrylamide to polymerize. Proteins are highly concentrated when they migrate through a stacking gel prior to entering a separating gel. The concentration occurs due to the difference in the rate of migration of glycine ion, chloride ion, and proteins, as illustrated below.

How to make acrylamide resolving gel?

In an Erlenmeyer flask, prepare the appropriate volume of solution containing the desired concentration of acrylamide for the resolving gel. Polymerization will begin as soon as the TEMED has been added. Without delay, swirl the mixture rapidly. Pour the acrylamide solution into the gap between the glass plates.

When was polyacrylamide gel used in electrophoresis?

Polyacrylamide gel (PAG) had been known as a potential embedding medium for sectioning tissues as early as 1964, and two independent groups employed PAG in electrophoresis in 1959. It possesses several electrophoretically desirable features that make it a versatile medium.

Where are molecular markers located in a polyacrylamide gel electrophoresis (PAGE)?

The molecular markers (ladder) are in the left lane Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility.

What acrylamide concentration should I use?

Use an appropriate gel concentration for your target protein. Using a higher acrylamide concentration produces a gel with a smaller mesh size suitable for the separation of small proteins. In general, an acrylamide concentration between 6 and 15% is used. Gels with an acrylamide concentration gradient (gradient gels) are also used.