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Mn

Zui Pan1, Sangyong Choi2, Yanhong Luo3

  • 1College of Nursing and Health Innovation, The University of Texas at Arlington, Arlington, TX, USA. zui.pan@uta.edu.

Methods in Molecular Biology (Clifton, N.J.)
|September 12, 2018
PubMed
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The manganese (Mn2+) quenching assay offers a straightforward method to measure store-operated calcium entry (SOCE) in cells. This technique utilizes manganese to quench Fura-2 fluorescence, providing a quantitative analysis of SOCE activity.

Area of Science:

  • Cellular Biology
  • Physiology
  • Biochemistry

Background:

  • Store-operated calcium entry (SOCE) is crucial for numerous cellular functions.
  • The fluorescent calcium indicator Fura-2 is commonly used to study SOCE.
  • Monitoring SOCE is essential for understanding cellular signaling.

Purpose of the Study:

  • To describe a Mn2+ quenching assay for measuring SOCE.
  • To detail the application of this assay in cultured esophageal epithelial and skeletal muscle cells.
  • To explain quantitative analysis of graded SOCE.

Main Methods:

  • Utilizing extracellular Mn2+ to enter cells via SOCE.
  • Measuring the fluorescence quenching of Fura-2 by Mn2+.
  • Applying the Mn2+ quenching assay to esophageal and skeletal muscle cells.
Keywords:
Esophageal squamous cell carcinomaFura-2-AMIsosbestic pointMn2+ quenchingRatiometric dyeSkeletal muscle myotubeStore-operated Ca2+ entry

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Main Results:

  • The Mn2+ quenching rate serves as a reliable indicator of SOCE extent.
  • This method allows for quantitative analysis of SOCE in different cell types.
  • The experimental method is effective for cultured esophageal epithelial and skeletal muscle cells.

Conclusions:

  • The Mn2+ quenching assay is a convenient and quantitative method for studying SOCE.
  • This technique facilitates the investigation of SOCE in various cellular contexts.
  • The described method aids in understanding the physiological roles of SOCE.