PlusOne N,N'-Methylene-bisacrylamide, 2% solution

How does %T and %C relate to my polyacrylamide gel?

The size of the pores in a polyacrylamide gel is determined by two parameters: total solids content (%T) and the ratio of cross-linker to acrylamide monomer (%C). The %T is the ratio of the sum of the weights of the acrylamide monomer and the cross-linker in the solution, expressed as % w/v. For example, a 20%T gel would contain 20% w/v of acrylamide plus bisacrylamide. As the %T increases, the pore size decreases. However, It has been found that for any single %T, 5% cross-linking creates the smallest pores in a gel. Above and below 5%, the pore size increases.

Which 2-D gel thickness should I use?

Either 1.0- or 1.5-mm-thick spacers can be used for all vertical formats. Thinner gels stain and destain more quickly and generally give less background staining. Thicker gels allow easier positioning of the IPG strip on the surface of the SDS gel and have higher protein capacity. Thicker gels are also less fragile and easier to handle.

How to avoid curved edges?

An alternative to using water-saturated butanol (to overlay the gels after casting) is to spray the cassettes using a 0.1 % (w/v) SDS/water solution (using a plant sprayer) such that the surface is covered by just a few millimeters. This technique helps to avoid curved edges on the gels.

Why did my polyacrylamide gel shrink a few centimeters after polymerization?

Too much catalyst used. Remake ammonium persulphate solution, to ensure correct concentration.