A smaller gel system for SDS-PAGE using tris-glycine buffer achieved clear separation of cardiac α,β-MHC isoforms and drastically reduced the total run time to 2 hours.
The study presents a faster (2 h) and efficient nongradient SDS-PAGE method for separating cardiac myosin heavy chain isoforms.
Abstract Current methods of SDS-PAGE for cardiac myosin heavy-chain (MHC) isoforms are complex and require more than 24 h. The aim of the current study was to improve the methodology of gel electrophoresis for faster and efficient separation of MHC isoforms. Rat ventricle and soleus tissues were subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) using tris-glycine buffer. Matrix-assisted laser desorption ionization (MALDI)–time of flight (TOF)–mass spectra (MS) of protein bands was done to identify α,β-MHC. Clear separation of α,β-MHC isoforms on SDS-PAGE was achieved and identity was confirmed by MALDI-TOF-MS. The smaller gel system drastically reduced the total run time to 2 h. The present method provides a simple, quick, and efficient protocol for cardiac α,β-MHC separation.
Kumar et al. (2009) studied this question. Smaller gel system for SDS-PAGE using tris-glycine buffer vs. Current methods (>24 h run time) was evaluated on Separation of α,β-MHC isoforms and total run time. A smaller gel system for SDS-PAGE using tris-glycine buffer achieved clear separation of cardiac α,β-MHC isoforms and drastically reduced the total run time to 2 hours.