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The CRAC Channel

Methods and Protocols

  • Book
  • © 2018

Overview

Editors:
  1. Aubin Penna

    1. IRSET Inserm UMR 1085/ UR1, Rennes, France

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    You can also search for this editor in PubMed   Google Scholar

  2. Bruno Constantin

    1. CNRS ERL 7003, Laboratoire STIM, Poitiers, France

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1843)

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Table of contents (16 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Patch-Clamp Recording of the CRAC Channel Current in STIM-Orai Overexpressing Cells

    • Yiming Zhang, Sisi Zheng, Yubin Zhou, Donald L. Gill, Youjun Wang
    Pages 1-16

  3. Fluorescence-Based Ratiometric Measurement of CRAC Channel Activity in STIM-Orai-Overexpressing HEK-293 Cells

    • Shuce Zhang, Lian He, Yubin Zhou, Youjun Wang
    Pages 17-39

  4. Recording SOCE Activity in Neurons by Patch-Clamp Electrophysiology and Microfluorometric Calcium Imaging

    • Hsiang-en Wu, Geza Gemes, Quinn H. Hogan
    Pages 41-53

  5. Mn2+ Quenching Assay for Store-Operated Calcium Entry

    • Zui Pan, Sangyong Choi, Yanhong Luo
    Pages 55-62

  6. Fluorescence-Based Measurements of Store-Operated Ca2+ Entry in Cancer Cells Using Fluo-4 and Confocal Live-Cell Imaging

    • Fujian Lu, Jianwei Sun, Tao Sun, Heping Cheng, Shengyu Yang
    Pages 63-68

  7. Fluorescence-Based Measurements of the CRAC Channel Activity in Cell Populations

    • Pedro C. Redondo, Alejandro Berna-Erro, Natalia Dionisio, Juan A. Rosado
    Pages 69-82

  8. Indirect Measurement of CRAC Channel Activity Using NFAT Nuclear Translocation by Flow Cytometry in Jurkat Cells

    • María D. Carretta, María A. Hidalgo, Rafael A. Burgos
    Pages 83-94

  9. CRAC Channel Components Quantitative Expression (In Tissues and Cell Lines) Using qPCR

    • Charlotte Dubois, Vyacheslav Lehenkyi, Natalia Prevarskaya
    Pages 95-106

  10. Western Blotting and Co-immunoprecipitation of Endogenous STIM/ORAI and Protein Partners

    • Charlotte Dubois, Natalia Prevarskaya
    Pages 107-113

  11. Study of Endogenous CRAC Channels in Human Mast Cells Using an Adenoviral Delivery System to Transduce Cells with Orai-Targeting shRNAs or with cDNAs Expressing Dominant-Negative Orai Channel Mutations

    • Ian Ashmole, Peter Bradding
    Pages 115-124

  12. Store-Operated Ca2+ Entry in Drosophila Primary Neuronal Cultures

    • Sumita Chakraborty, Gaiti Hasan
    Pages 125-136

  13. Study of the Endogenous CRAC Channel Using shRNA-Mediated Gene Silencing

    • Xuexin Zhang, Amy M. Spinelli, Wei Zhang, Mohamed Trebak
    Pages 137-145

  14. Engineered Cross-Linking to Study the Pore Architecture of the CRAC Channel

    • Guolin Ma, Lian He, Ji Jing, Peng Tan, Yun Huang, Yubin Zhou
    Pages 147-166

  15. Measurement of the CRAC Channel Fast Ca2+-Dependent Inactivation (FCDI)

    • Grigori Y. Rychkov
    Pages 167-173

  16. High-Resolution Imaging of STIM/Orai Subcellular Localization Using Array Confocal Laser Scanning Microscopy

    • Andras T. Deak, Benjamin Gottschalk, Emrah Eroglu, Rene Rost, Markus Waldeck-Weiermair, Wolfgang F. Graier et al.
    Pages 175-187

  17. Single-Channel Single-Molecule Detection (SC-SMD) System

    • Laura G. Ceballos, Alexander Asanov, Luis Vaca
    Pages 189-201

  18. Back Matter

    Pages 203-204
    Download chapter PDF
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Keywords

About this book

This volume discusses the cellular and molecular techniques used to study and characterize the different components of the CRAC channel signaling pathway. The chapters in this book cover topics such as fluorescence-based measurements of the CRAC channel activity in cell populations; patch-clamp recording of the CRAC channel in STIM-Orai overexpressing cells and native systems; western-blotting and co-immunoprecipitation of endogenous STIM/ORAI and protein partners; shRNA-mediated gene silencing; and the SC-SMD system. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.



Thorough and cutting-edge, The CRAC Channel: Methods and Protocols is a valuable resource for any researcher interested in learningmore about the CRAC channel, as its activation mechanism and roles in cellular functions have yet to be completely explored.


Editors and Affiliations

  • IRSET Inserm UMR 1085/ UR1, Rennes, France

    Aubin Penna

  • CNRS ERL 7003, Laboratoire STIM, Poitiers, France

    Bruno Constantin

Bibliographic Information

  • Book Title: The CRAC Channel

  • Book Subtitle: Methods and Protocols

  • Editors: Aubin Penna, Bruno Constantin

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-8704-7

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2018

  • Hardcover ISBN: 978-1-4939-8702-3Published: 10 September 2018

  • Softcover ISBN: 978-1-4939-9370-3Published: 10 December 2019

  • eBook ISBN: 978-1-4939-8704-7Published: 10 September 2018

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 204

  • Number of Illustrations: 15 b/w illustrations, 26 illustrations in colour

  • Topics: Biochemistry, general

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