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Fura-2 Calcium Assay Kit - No Wash (ab176766) is a ratiometric calcium assay that allows homogeneous measurement of intracellular calcium mobilization caused by activation of G-protein-coupled receptors (GPCR) or calcium channels. Cells expressing a GPCR of interest that signals through calcium are pre-loaded with Fura-2 AM which can cross cell membrane. Once inside the cell, the lipophilic blocking groups of Fura-2 AM are cleaved by esterases, resulting in a negatively charged fluorescent dye that stays inside cells and its fluorescence wavelength is blue-shifted upon binding to calcium. When cells are stimulated with agonists, the receptor signals the release of intracellular calcium, which greatly increase the fluorescence intensity of Fura-2 at the short wavelength.
The characteristics of its ratio measurements reduce the effects of uneven dye loading and cell numbers, dye leakage and photo bleaching. The 340/380 nm excitation ratio allows accurate measurements of the intracellular Ca2+ concentration and makes this kit an ideal tool for more accurate measurement of cellular calcium concentration compared to Fluo-8 of the single wavelength.
ab176766 provides an optimized assay method with no washing step for monitoring the G-protein-coupled receptors and calcium channels using HTS instrumentation. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format.
This product is intended to be used for monitoring calcium fluctuations in vivo in live cells using the following HTS imaging plate readers: FLIPR™, FDSS, BMG NOVOstar™, FLexStation, ViewLux, IN Cell Analyzer or Arrayscan.
|Components||10 x 96 tests||100 x 96 tests|
|10X Pluronic® F127 Plus||10 x 1ml||10 x 10ml|
|Fura-2 AM||1 vial||10 vials|
|HHBS||1 x 100ml||0 x 0ml|
CHO-K1 cells were seeded overnight at 40,000 cells/100 μL/well in a Costar black wall/clear bottom 96-well plate. The cells were incubated with 100 μL of the Fura-2 No Wash Calcium Assay Kit (ab176766) for 1 hour at room temperature. ATP (50 μL/well) was added to achieve the final indicated concentrations.
ab176766 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"