Measurement of phosphorylated phospholamban levels in cardiomyocytes (HL-1) by immunoprecipitation.

C. V. Suresh Babu; Eun Joo Song; Sheikh Md Enayetul Babar; Young Sook Yoo

doi:10.1016/j.jbbm.2007.05.008

Measurement of phosphorylated phospholamban levels in cardiomyocytes (HL-1) by immunoprecipitation.

C. V. Suresh Babu, Eun Joo Song, Sheikh Md Enayetul Babar, Young Sook Yoo

Pharmaceutical Sciences

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Abstract

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.

Original language	English
Pages (from-to)	838-844
Number of pages	7
Journal	Journal of Biochemical and Biophysical Methods
Volume	70
Issue number	6
DOIs	https://doi.org/10.1016/j.jbbm.2007.05.008
State	Published - 24 Apr 2008

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@article{52571cb197f64cc988271ff368dc03f1,

title = "Measurement of phosphorylated phospholamban levels in cardiomyocytes (HL-1) by immunoprecipitation.",

abstract = "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.",

author = "{Suresh Babu}, {C. V.} and Song, {Eun Joo} and Babar, {Sheikh Md Enayetul} and Yoo, {Young Sook}",

note = "Funding Information: The authors wish to thank Mrs Anne J. Charlton for English proof reading and helpful comments on the manuscript. This work was supported by the grants from the Korea Ministry of Science and Technology (Systems Biology Research Grant; 2N29550).",

year = "2008",

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doi = "10.1016/j.jbbm.2007.05.008",

language = "English",

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T1 - Measurement of phosphorylated phospholamban levels in cardiomyocytes (HL-1) by immunoprecipitation.

AU - Suresh Babu, C. V.

AU - Song, Eun Joo

AU - Babar, Sheikh Md Enayetul

AU - Yoo, Young Sook

N1 - Funding Information: The authors wish to thank Mrs Anne J. Charlton for English proof reading and helpful comments on the manuscript. This work was supported by the grants from the Korea Ministry of Science and Technology (Systems Biology Research Grant; 2N29550).

PY - 2008/4/24

Y1 - 2008/4/24

N2 - 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.

AB - 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.

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U2 - 10.1016/j.jbbm.2007.05.008

DO - 10.1016/j.jbbm.2007.05.008

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AN - SCOPUS:48649094069

SN - 0165-022X

VL - 70

SP - 838

EP - 844

JO - Journal of Biochemical and Biophysical Methods

JF - Journal of Biochemical and Biophysical Methods

IS - 6

ER -

Measurement of phosphorylated phospholamban levels in cardiomyocytes (HL-1) by immunoprecipitation.

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