CAS:130100-20-8
Storage:Powder:-20℃,2 years;Insolvent(Mother Liquid):-20℃,6 months;-80℃,1 year
Purity:≥95%
Appearance:Solid
Mag-Fura-2 AM is an intracellular magnesium ion indicator, which can also be used to detect calcium ion concentration, and is a ratio probe excited by ultraviolet. Similar to Fura-2, the excitation wavelength of Mag-Fura-2 undergoes a blue transition from 369nm to 330nm. Mag-Fura-2 AM is permeable to cell membranes and can be loaded into cells after simple incubation.
Fluorescence microscope (for reference only)
Excitation Fura 2 filter set
Emission Fura 2 filter set
Recommended plate Black wall/clear bottom
Fluorescence microplate reader (for reference only)
Excitation 340, 380
Emission 510
Cutoff 475
Recommended plate Black wall/clear bottom
Instrument specification(s) Bottom read mode/Programmable liquid handling
Example Protocol (for reference only)
Mag-Fura-2 AM Working Solution
For most cell lines, Mag-Fura-2 AM at a final concentration of 4 to 5 μM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.
If it is necessary to prepare a stock solution, it is recommended to store in aliquots to avoid product failure caused by repeated freezing and thawing.
Note If your cells contain organic anion-transporters, probenecid (1-2 mM) may be added to the dye working solution (final in well concentration will be 0.5-1 mM) to reduce leakage of the de-esterified indicators.
SAMPLE EXPERIMENTAL Protocol (for reference only)
Following is our recommended protocol for loading AM esters into live cells. This protocol only provides a guideline and should be modified according to your specific needs.
Prepare cells in growth medium overnight.
On the next day, add 1X Mag-Fura-2 AM working solution into your cell plate.
Note If your compound(s) interfere with the serum, replace the growth medium with fresh HHBS buffer before dye-loading.
Incubate the dye-loaded plate in a cell incubator at 37 ℃ for 30 to 60 minutes.
Note Incubating the dye for longer than 1 hour can improve signal intensities in certain cell lines.
Replace the dye working solution with HHBS or buffer of your choice (containing an anion transporter inhibitor, such as 1 mM probenecid, if applicable) to remove any excess probes.
Add the stimulant as desired and simultaneously measure fluorescence using either a fluorescence microscope equipped with a Fura 2 filter set or a fluorescence plate reader containing a programmable liquid handling system such as a FlexStation, at Ex/Em1 = 340/510 nm cutoff 475 nm and Ex/Em2 = 380/510 nm cutoff 475 nm.