Electrophoresis is a popular technique used to separate closely related compounds. It is based on the movement of charged particles in the electric field. This technique is based on the principle that positively charged particles migrate toward cathode and negatively charged particles migrate toward the anode. Rate of migration depends on the charge of the molecule. Application of electrophoresis, factors affecting migration of charged particles, types of electrophoresis, basic requirements of electrophoresis, paper electrophoresis, separation of plasma proteins, and movement of different protein fractions are covered in this chapter.