Human macrophages were loaded with 1.25 μm of Fluo3-AM (Thermo Fisher Scientific) for 30 min at room temperature in loading buffer containing 150 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 20 mM Hepes, 10 mM glucose, 1× PowerLoad (Thermo Fisher Scientific) to solubilize the Fluo3-AM dye, and 2.5 mM probenecid (Thermo Fisher Scientific) for dye retention. Fluo3-AM was then removed, and the cells were chased in loading buffer without PowerLoad and probenecid for 30 min at room temperature. Cytosolic Ca2+ was monitored by flow cytometry (FACSCanto II) or with a Nikon A1 confocal microscope.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.