Sep 05, 2022

Public workspaceFluo-3-AM Assay

This protocol is a draft, published without a DOI.
  • 1Fluo Calcium Indicators
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Protocol Citation: Thermo Fisher 2022. Fluo-3-AM Assay. protocols.io https://protocols.io/view/fluo-3-am-assay-cf6qtrdw
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: September 04, 2022
Last Modified: September 05, 2022
Protocol Integer ID: 69552
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Abstract
determine the intracellular Ca2+ concentration of your cells via a fluorescence marker
dilute the DMSO stock solution (1-5mM) to 1-5μM in the buffered physiological medium of choice
Pluronic F-127 may be added to a concentration of Concentration0.02 % volume

mix in an aliquot of Fluo-3-AM stock solution to a concentration of Concentration4 micromolar (µM)

incubate an aliquot of the cells with Fluo-3-AM for Duration01:00:00 at Temperature20 °C -Temperature37 °C
1h
wash the cells in indicator-free medium and incubate the cells for another Duration00:30:00 in dye-free medium

30m
measure fluorescence at Ex/Em = 506/526 with a microplate reader