Measurement of phosphorylated phospholamban levels in cardiomyocytes (HL-1) by immunoprecipitation.
Phospholamban (PLN) is a key regulatory protein involved in cardiac calcium signaling through the pumping of cytoplasmic Ca2+ into the sarcoplasmic reticulum (SR). Recent systems-level studies have focused on integrating quantitative data (e.g. protein expression levels) for a better understanding of cardiac systems biology. In this view, we developed a capillary electrophoresis (CE) based immunoprecipitation method for the measurement of phospho-PLN (ser 16) in cardiomyocytes (HL-1 cell line). Dose-dependent isoproterenol (Iso) treated cells were analyzed using CE, and the phospho-PLN levels were quantified using specific polyclonal antibodies. The CE method employed was accurate, quick and easier compare to other techniques and the results are useful for the subsequent computational systems biology research.
Suresh Babu
C V
CV
Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology, P.O. BOX 131, Cheongryang, Seoul, 130-650, Republic of Korea.
Song
Eun Joo
EJ
Babar
Sheikh Md Enayetul
SM
Yoo
Young Sook
YS
D002135:Calcium-Binding Proteins / Q000032:analysis* / Q000378:metabolism; D002460:Cell Line; D019075:Electrophoresis, Capillary; D047468:Immunoprecipitation / Q000379:methods*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D000099211:Phospholamban
D002135:Calcium-Binding Proteins; D000099211:Phospholamban