Purification of Myosin Heavy Chain Isoforms by Electroendos- motic Preparative Gel Electrophoresis: Characterization of Em- bryonic Slow Myosin Heavy Chain

Myosin is a hexapolypeptide constituted by four light and two heavy chains the isoforms of which segregate differently in specific stages of animal development and in different fiber types in adulthood. In mammals the Myosin Heavy Chains (MHC) are polypeptides with a molecular mass of about 200 KDa which isoforms can be identified by SDS PAGE and/or immunochemistry. A method for the purification of myosin heavy chain isoforms using a SDS Electrendosmotic Preparative Gel Electrophoresis (SDS EPGE) is described. Semplicity and reproducibility of this approach permits purification of single isoforms from complex mixture with a sufficient high recovery to perform immunochemical or biochemical studies. The possibility to apply useful tool as SDS removal and protein concentration by KDS precipitation before further analysis on the sample is described. Application of the methodology to the study of slow type embryonic MHC by enzymatic as well as chemical peptide mapping and amino acid composition is described.