Along with his preliminary results, Itano suggested in a 1948 report that there were eight other options for immediate research, one of which eventually proved successful: "electrophoretic studies on normal and sickle cell hemoglobin." In his synopsis about electrophoresis, Itano briefly mentioned the preliminary status of electrophoretic studies on hemoglobin and its possible utility for his research. Itano stated later that he initially tried electrophoresis because the apparatus was available and required small amounts of blood. At the time, Caltech had a problem getting adequate samples of blood from people with sickle cell anemia. Pauling finally obtained a regular supply in May 1949 from Dr. George Burch of Tulane University.

Electrophoresis uses electric currents to examine substances that have varying surface charges, and therefore is especially helpful in analyzing proteins. Protein samples in a buffer solution are run through the electrophoresis apparatus, which creates a moving boundary. Moving boundary electrophoresis indicates whether a protein has an overall positive or negative charge on its surface area. The rate of a moving boundary can be manipulated by changing the pH of the solution and hence, the charge of the particles.

Since moving boundary electrophoresis was a relatively new technique in 1948, only about fifty laboratories in the United States had electrophoresis machines. Mass production of the apparatus began after 1950, so that prior to mid-century, laboratories built them. Pauling asked the Rockefeller Foundation in 1941 for money for immunology research, including the construction of a Tiselius electrophoresis apparatus. Caltech received enough money from the Rockefeller Foundation to build the machine, to which Stanley M. Swingle, a general chemistry instructor at Caltech, made improvements. It was this machine that Itano and Seymour Jonathan Singer used for the experiments reported in the 1949 article.

Pauling's request for money to build an electrophoresis machine was most likely influenced by Karl Landsteiner's infatuation with the device. Landsteiner, Pauling's mentor in immunology, analyzed blood sera with electrophoresis beginning in 1908. When Landsteiner learned of Arne Tiselius's improved apparatus in 1937 he immediately began using the technique as a standard procedure in his researches.