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APS and TEMED are the polymerization starting components basically used in the polyacrylamide gel electrophoresis. During the polyacrylamide gel electrophoresis, acrylamide and bisacrylamide are added to form a gel matrix. Acrylamide forms polymers in which bisacrylamide creates a cross-linking. Thus, it gives the overall gel-like matrix with varying pore size depending upon the concentration of bisacrylamide.
APS or ammonium persulfate is the one which initiates polymerization of the acrylamide and bisacrylamide. TEMED, the full name of which is (N,N,N’,N’ Tetramethylethylene-1,2-diamine is added at the last in the gel mixture. It reacts with the ammonium persulfate and causes splitting of the persulfate ions into sulfate free radicals. Thus formed sulfate free radicals now initiate the polymerization (free radical reaction) of the acrylamide and cross-linking with bisacrylamide.
An electrophoresis is an analytical tool used in the biochemistry laboratory to separate and characterize proteins and nucleic acids based on their net molecular charge to mass ratio. There are different types of electrophoresis like capillary electrophoresis, agarose gel electrophoresis, polyacrylamide gel electrophoresis, etc. Polyacrylamide gel electrophoresis can be of two types; denaturing and non-denaturing.
SDS-PAGE is a denaturing polyacrylamide gel electrophoresis, frequently used in the biochemistry laboratory to separate and characterize proteins. In this type of electrophoresis, sodium dodecyl sulfate (SDS) is used to denature the protein and provide equal mass to charge ratio so as to allow the protein to move constantly through the gel based on their molecular weight. In SDS-PAGE, 2-mercaptoethanol is also added to cleave the disulfide linkage if any present.