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GFP-CERTIFIED® FLUOFORTE® Calcium assay kit

A brighter, more robust and GFP-compatible (orange dye) fluorescent assay for calcium mobilization
 
ENZ-51020-KP010 1 Kit 444.00 USD
 
ENZ-51020-KP100 1 Kit 3,188.00 USD
Do you need bulk/larger quantities?
 
  • Monitor calcium mobilization in GFP-expressing cell lines
  • The brightest and most sensitive red-shifted fluorescent Ca2+ indicator
  • Provides comparable EC50 and IC50 values as obtained using Rhod-4 AM and Fluo-4 AM
  • Larger assay window and higher Z' factor values, allowing measurements of challenging cell lines and receptors
Enzo Life Sciences’ GFP-CERTIFIED® FLUOFORTE® Calcium Assay Kit (high-throughput) provides a homogeneous fluorescence-based assay for detecting intracellular calcium mobilization across a broad spectrum of biological targets. Relative to other commercially available dyes, the GFP-CERTIFIED® FLUOFORTE® dye is the brightest and most sensitive fluorescent calcium indicator. The kit provides a homogeneous mix-and-read, no-wash calcium mobilization assay. The homogenous cell-based assay for calcium offers fewer steps, lower variability and an easier protocol for adherent and non-adherent cell lines. In addition, it requires neither a washing step, nor exogenous addition of a quencher dye, which could adversely effect receptor-ligand interaction kinetics.
ENZ-51020 Fig3
Histamine H1 receptors: HeLa cells were seeded overnight at 40,000 cells per 100 µL per well in a 96-well black wall/clear bottom microplate. The cells were incubated with 100 µL of GFP-CERTIFIED® FLUOFORTE® reagent or Rhod-2 AM dye for 1 hour at 37°C. Histamine (20µL/well) was added using a BioTek two-syringe pump dispenser to achieve the final indicated concentrations. No significant difference in EC50 of Histamine for GFP-CERTIFIED® FLUOFORTE® and Rhod2-AM dye was observed. GFP-CERTIFIED® FLUOFORTE® reagent generated higher intensity signal and a larger assay window.
ENZ-51020 FLUOFORTE vs RHOD-2
U-2 OS cells were incubated with (A) GFP-CERTIFIED® FLUOFORTE® AM dye or (B) Rhod-2 AM dye. Cells were washed and treated with a final concentration of 100nM ATP and imaged immediately. GFP-CERTIFIED® FLUOFORTE® AM dye exhibits brighter fluorescence intensity than Rhod-2 AM.
ENZ 51020 spectra
Please mouse over
ENZ-51020 Fig3 ENZ-51020 FLUOFORTE vs RHOD-2 ENZ 51020 spectra

Product Specification

Applications:Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS
 
Application Notes:The GFP-CERTIFIED® FLUOFORTE® Calcium Assay kit provides a homogeneous fluorescence-based assay for detecting intracellular calcium mobilization across a broad spectrum of biological targets.
 
Quality Control:% purity of Reagent A (GFP-CERTIFIED® FLUOFORTE® dye) by HPLC is ≥95%.
 
Quantity:ENZ-51020-KP010: for 10 x 96-well plates
ENZ-51020-KP100: for 100 x 96-well plates
 
Handling:Protect from light. Avoid freeze/thaw cycles.
 
Shipping:Shipped on Dry Ice
 
Short Term Storage:-20°C
 
Long Term Storage:-80°C
 
Kit/Set Contains:Starter pack (Prod. No. ENZ-51020-KP010):
Reagent A (lyophilized GFP-CERTIFIED® FLUOFORTE® dye), 1 vial
Reagent B (dye efflux inhibitor), 10ml
Reagent C (Hanks’ buffer with 20mM HEPES, 100ml)
Sufficient for 10 x 96-well plates

High-throughput (Prod. No. ENZ-51020-KP100):
Reagent A (lyophilized GFP-CERTIFIED® FLUOFORTE® dye), 10 vials
Reagent B (dye efflux inhibitor), 10 x 10ml
Sufficient for 100 x 96-well plates
 
Technical Info/Product Notes:The GFP-CERTIFIED® FLUOFORTE® Calcium assay kit is a member of the CELLESTIAL® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications. CELLESTIAL® reagents and kits are optimal for use in demanding cell analysis applications involving confocal microscopy, flow cytometry, microplate readers and HCS/HTS, where consistency and reproducibility are required.
 

Product Literature References

Gα13/PDZ-RhoGEF/RhoA signaling is essential for gastrin-releasing peptide receptor-mediated colon cancer cell migration: M. Patel, et al.; Mol. Pharmacol. 86, 252 (2014), Application(s): Microplate reader using Caco-2 cells, Abstract; Full Text
The N-terminally truncated µ3 and µ3-like opioid receptors are transcribed from a novel promoter upstream of exon 2 in the human OPRM1 gene: S. Andersen, et al.; PLoS One. 8, e71024 (2013), Application(s): Microplate reader using HEK293 cells, Abstract; Full Text
Leukotriene C4 induces migration of human monocyte-derived dendritic cells without loss of immunostimulatory function: J. Dannull, et al.; Blood 119, 3113 (2012), Application(s): Intracellular calcium release in human monocyte derived dendritic cells with Tecan Infinite plate reader, Abstract; Full Text

General Literature References

Novel Fluo-4 analogs for fluorescent calcium measurements: V.V. Martin, et al. ; Cell Calcium 36, 509 (2004), Abstract;
Calcium measurements in perfused mouse heart: quantitating fluorescence and absorbance of Rhod-2 by application of photon migration theory: C. Du, et al.; Biophys. J. 80, 549 (2001), Full Text
Rhod-2 based measurements of intracellular calcium in the perfused mouse heart: cellular andsubcellular localization and response to positive inotropy: G.A. MacGowan, et al.; J. Biomed. Opt. 6, 23 (2001), Abstract;
Flow cytometric kinetic assay of calcium mobilization in whole blood platelets using Fluo-3 and CD41: M. do Céu Monteiro, et al.; Cytometry 35, 302 (1999), Abstract;
Nucleoplasmic and cytoplasmic differences in the fluorescence properties of the calcium indicator Fluo-3: C. Perez-Terzic, et al.; Cell Calcium 21, 275 (1997), Abstract;
Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets: R. Greimers, et al.; Cytometry 23, 205 (1996), Abstract;

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