BCHM 426 Lecture Notes - Lecture 3: Polyacrylamide Gel Electrophoresis, Free-Radical Reaction, Acrylamide

• School: Binghamton University
• Department: Biochemistry
• Course: BCHM 426
• Professor: mark

Document Summary

Polyacrylamide gel electrophoresis and transfer to pvdf membrane. Run complex mix of proteins in gel matrix to separate proteins based on their properties. If we run a standard, we calculate the migration of each protein. If it is set up where negative ions run to positive anode, some may run in the opposite direction. Separates proteins so electrophoresis can also serve as a first step in immunochemical techniques such as western blot. Protein molecules have a net charge when the ph isnt equal to pi. This is because there are ionizable groups in the amino acid. Have at least 2 residues that can be ionized, such as amino and carboxy groups. Thus, the more ionizable groups there are, the more negative charges there are. Every protein has a different pi, so they may have different charges. They wont move in a consistent way if all you did was use the intrinsic charge of the proteins.