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Related Experiment Video

Updated: May 10, 2026

Monitoring ER/SR Calcium Release with the Targeted Ca2+ Sensor CatchER+
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Measuring Ca2+ release evoked by cyclic adp-ribose.

Andreas H Guse1

  • 1The Calcium Signalling Group, Department of Biochemistry and Signal Transduction and Department of Biochemistry and Molecular Cell Biology, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany. guse@uke.de

Cold Spring Harbor Protocols
|June 5, 2013
PubMed
Summary
This summary is machine-generated.

Measuring calcium (Ca2+) release evoked by cyclic ADP-ribose (cADPR) is challenging. This study details step-by-step methods for delivering cADPR to cells using permeabilization or microinjection.

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Area of Science:

  • Cellular Biology
  • Biochemistry
  • Physiology

Background:

  • Cyclic ADP-ribose (cADPR) is a ubiquitous second messenger involved in calcium (Ca2+) mobilization.
  • Measuring cADPR-evoked Ca2+ release presents practical difficulties due to cADPR's inability to cross cell membranes at physiological pH.

Purpose of the Study:

  • To provide detailed, step-by-step protocols for delivering cADPR into the cytosol for Ca2+ release studies.
  • To address the challenges associated with measuring cADPR activity in various cell types.

Main Methods:

  • The study describes two primary methods: permeabilizing the plasma membrane and microinjection.
  • Protocols are specifically optimized for the Jurkat T-lymphoma cell line.
  • Adaptations for other cell types are acknowledged.

Main Results:

  • Successful delivery of cADPR into the cytosol was achieved using both permeabilization and microinjection techniques.
  • These methods facilitate the measurement of Ca2+ release evoked by cADPR.
  • The described protocols enable robust investigation of cADPR signaling pathways.

Conclusions:

  • The provided methods offer practical solutions for overcoming challenges in studying cADPR-mediated Ca2+ signaling.
  • These techniques are crucial for understanding the physiological roles of cADPR in different cell types.
  • Further adaptation may be necessary for cell types beyond Jurkat T-lymphoma cells.