Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>

How do you measure Ca2+ response with calcium indicators like Calbryte 520, potassium salt?

Posted April 1, 2019


Answer

Below is a sample protocol for measuring calcium response using Calbryte™ 520, potassium salt (Cat# 20658).

  1. Begin by creating 10-15 µM probe solution in the following buffer: 
    • 10 mM K2EGTA in 100 mM KCl and 30 mM MOPS, pH 7.2:  Free Ca2+ in this solution (20o C) ~0 µM
    • 10 mM Ca2+ + 10mM EGTA in 100 mM KCl and 30 mM MOPS, pH 7.2:  Free Ca2+ in this solution (20o C) ~39 µM 
  1. Measure the fluorescence change at Ex/Em= 490/525 nm (cutoff=515 nm) with a microplate reader or fluorescence spectrometer.
  2. Calculate the fold of intensity increase.
Additional resources

Calbryte-520 Potassium Salt