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Detection of Calcium Binding Proteins on Nitrocellulose Membrane

Research Project

Project/Area Number 57880034
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field 分子遺伝学・分子生理学
Research InstitutionThe University of Tokyo
Okazaki National Research Institutes

Principal Investigator

MIKAWA Takashi  岡山国立共同研究機構, その他 (50143417)

Project Period (FY) 1982 – 1985
Project Status Completed (Fiscal Year 1985)
Budget Amount *help
¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1983: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1982: ¥3,400,000 (Direct Cost: ¥3,400,000)
KeywordsCalcium Binding Protein / アフィニティ分析 / ウエスタンブロット
Research Abstract

A new simple method of detecting calcium binding proteins in a protein mixture is described. A sample which might include calcium binding proteins was subjected to SDS-polyacrylamide gel electrophoresis and then electrophoretically transferred to a nitrocellulose membrane. The membrane was then incubated with <^(45)Ca> to detect calcium binding proteins as radioactive bands by autoradiography.
Purified troponin-C, calmodulin, myosin DTNB light chain, and parvalbumin were clearly identified by this method. In the whole homogenate of chicken skeletal muscle, myosin DTNB light chain, troponin-C, and 55K calcium binding protein were found to be radioactive. In the frog skeletal muscle, small molecular weight proteins of approximately 13-15K and 70K protein appeared to be the calcium binding proteins. In the case of the carp skeletal muscle, small molecular weight proteins including parvalbumin and two proteins of about 80K seemed to bind calcium ion. Two high molecular weight calcium binding proteins were present in the scallop striated muscle.
The procedure described can be completed within 24 h and can detect as little as 2 <micro> g of calcium binding protein in the starting sample. Under appropriate conditions it was possible to detect only high affinity calcium binding proteins.

Report

(1 results)
  • 1985 Final Research Report Summary
  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] J.Biochem. 93-3. (1983)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J.Biochem. 94-3. (1983)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J.Biochem. 95-2. (1984)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1985 Final Research Report Summary
  • [Publications] J. Biochem. 95-2. (1984)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1985 Final Research Report Summary

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Published: 1987-03-31   Modified: 2016-04-21  

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