What is gradient polyacrylamide gel electrophoresis?

Gradient polyacrylamide gel electrophoresis is a powerful tool for the resolution of polypeptides by relative mobility. Here, we present a simplified method for generating polyacrylamide gradient gels for routine analysis without the need for specialized mixing equipment.

What is polyacrylamide gel electrophoresis (PAGE)?

Part of the book series: Methods in Molecular Biology ( (MIMB,volume 869)) Polyacrylamide gel electrophoresis (PAGE) is one of the most powerful tools used for protein analysis. We describe the use of Tris–acetate buffer and 3–15% polyacrylamide gradient gels to simultaneously separate proteins in the mass range of 10–500 kDa.

What is protein electrophoresis in a gel matrix?

Electrophoresis in a gel matrix stands as the most popular technique, employing a variety of gel materials such as starch gels, dextran gels, agarose gels, and polyacrylamide gels . In this protocol, we will focus on protein electrophoresis in polyacrylamide gels.

What materials are used for electrophoresis?

Electrophoresis can be carried out in free solutions (e.g., capillary electrophoresis) or with supporting media such as papers, films, and gels. Electrophoresis in a gel matrix stands as the most popular technique, employing a variety of gel materials such as starch gels, dextran gels, agarose gels, and polyacrylamide gels .

Which acrylamide concentration is best for electrophoresis?

While the acrylamide concentration can be altered to produce different migration patterns, electrophoresis with a gel of fixed concentration offers optimal resolution and resolving power within a limited range of polypeptide masses . Therefore, it is often desirable to use gels with an acrylamide concentration gradient.

Do gradient acrylamide gels increase pore size?

However, the use of polyacrylamide gels that have a gradient of increasing acrylamide concentration (and hence decreasing pore size) can sometimes have advantages over fixed-concentration acrylamide gels. During electrophoresis in gradient gels, proteins migrate until the decreasing pore size impedes further progress.