What is fluo 8?

What is fluo 8?

Fluo-8® is a novel green calcium indicator which incorporates the same fluorescein core, utilized in Fluo-3 and Fluo-4, to monitor Ca2+ concentration and flux in cells.

How do you dissolve fluo-4 am?

Dissolve Fluo-4 AM in 25 µl of DMSO. After adding DMSO, vortex tube briefly to dissolve the indicator dye, then centrifuge briefly to collect all contents at the tube bottom. Add entire contents of indicator dye tube to assay buffer solution to make a dye loading solution.

How does fluo-4 AM work?

Fluo-4 is an analog of Fluo-3 with the two chlorine substituents replaced by fluorines, which results in increased fluorescence excitation at 488 nm that gives higher fluorescence signal. Fluo-4 has its absorption maximum at 494 nm, thus making it excitable by the argon-ion laser.

How does Fura 2 work?

Fura-2, an aminopolycarboxylic acid, is a ratiometric fluorescent dye which binds to free intracellular calcium. The use of the ratio automatically cancels out confounding variables, such as variable dye concentration and cell thickness, making Fura-2 one of the most appreciated tools to quantify calcium levels.

How is calcium influx measured?

There are several directly or indirectly ways to measure intracellular Ca2+ influx:

  1. Using Ca2+ sensitive dye such as FURa-2 as recommended to you by others.
  2. using intracellular Ca2+ sensitive microelectrode.
  3. Performing Whole cell patch clamp recording.

What is calcium flux?

Indo-1 is the ratio metric calcium indicator dye most commonly used in flow cytometry because of its shift in emission frequency when excited at a single wavelength. …

Why is Fura-2 unable to cross cell membranes?

Fluorescent Dyes – Loading Calcium indicators are unable to cross lipid membranes due to their nature, making necessary the use of physical or chemical methods to load them inside the cell. This is the case of Indo-1 AM and Fura-2 AM, which are fluorescent and calcium insensitive.

What can fluo-8 am be used for?

Fluo-8® AM has been widely used to study calcium ions in critical biological processes across a span of different disciplines. Such processes include, but are not limited to, G protein-coupled receptor signaling pathways, calcium ion channel activity, intracellular/cytosolic Ca2+ flux and activation of cell receptors.

What should be the final concentration of fluo-8 am?

For most cell lines, Fluo-8® AM at a final concentration of 4-5 μM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.

Which is better for calcium binding Fluo 4 or Fluo 8?

Binds to intracellular calcium (K d = 390 nM). Fluorescence intensity increases upon Ca 2+ binding. Cell-permeable. Fluo-8 (or Fluo-2 Medium Affinity) has been found to be brighter (1.5x) than Fluo-4 in cellular experiments.

Which is brighter Fluo 4 am or Fluo 8 am?

In addition, Fluo-8® AM is two times brighter than Fluo-4 AM and four times brighter than Fluo-3 AM. AAT Bioquest offers a set of our outstanding Fluo-8® reagents with different calcium-binding affinities (Fluo-8® Kd = 389 nM; Fluo-8H™ Kd = 232 nM; Fluo-8L™ Kd = 1.86 µM; Fluo-8FF™ Kd = 10 µM).

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