Experimental Set-up for Poly Acrylamide Gel Electrophoresis

As explained above a solid support during electrophoresis helps in separation of molecules in process of electrophoresis. A very common method for separating proteins by electrophoresis uses a polyacrylamide gel as a support medium. The process is called polyacrylamide gel electrophoresis. Polyacrylamide gels are formed from the polymerization of two compounds, acrylamide and N,N ' -methylene- bis-acrylamide. N,N'-methylene- bis-acrylamide is a cross-linking agent for the gels. The polymerization is initiated by the addition of freshly prepared ammonium persulfate along with N,N,N¹,N¹,- tetramethylethylenediamine (TEMED). The TEMED has ability to exist as a free radical and acts as an additional catalyst for the polymerization (Fig. 1). The polymerization is done in gel polymerized between two plates of glass. Details experimental set-up is shown in series of figures (Fig.2 A-C).

Figure 1: Polymerization reaction of acrylamide and bis-acrylamide