How does polyacrylamide gel electrophoresis (PAGE) work?

Gel electrophoresis is a fundamental technique for separating molecules such as DNA, RNA and proteins in laboratories across the biological disciplines. In this article, we will consider how polyacrylamide gel electrophoresis (PAGE) works, how it can be interpreted and some of its applications.

What is polyacrylamide gel electrophoresis (SDS-PAGE)?

The most commonly used form of polyacrylamide gel electrophoresis is the Sodium dodecyl suplhate Polyacrylamide gel electrophoresis (SDS- PAGE) used mostly for the separation of proteins. SDS-PAGE (Polyacrylamide Gel Electrophoresis), is an analytical method used to separate components of a protein mixture based on their size.

What does %t mean in polyacrylamide gel electrophoresis?

%T indicates the pore size of the prepared gel. Polyacrylamide gel electrophoresis is based on the principle that charged particles migrate to the electrode of the opposite sign under the influence of an electric field. Principle of polyacrylamide gel electrophoresis, Image source: DOI: 10.1016/B978-0-12-803077-6.00012-6

Why is two-dimensional polyacrylamide gel electrophoresis important?

In cases such as this, separation in two dimensions can add the required resolving power as it is less likely that two molecules will be very similar in two distinct properties. Two-dimensional polyacrylamide gel electrophoresis, or 2D PAGE, was introduced in 1975 concurrently by Joachim Klose 7 and Patrick H. O’Farrel. 8

How does a polyacrylamide gel separate analytes?

The basic principle of PAGE is to separate analytes by passing them through the pores of a polyacrylamide gel using an electric current. To achieve this, an acrylamide– bisacrylamide mix is polymerized (polyacrylamide) by the addition of ammonium persulfate (APS).

What is a general electrophoresis technique?

The technique is based upon the principle that a charged molecule will migrate in an electric field towards an electrode with opposite sign. The general electrophoresis techniques cannot be used to determine the molecular weight of biological molecules because the mobility of a substance in the gel depends on both charge and size.