Search
Citations & References (21)
Invitrogen™
Fluo-4 NW Calcium Assay Kit
Fluo-4 NW (No-Wash) Calcium Assay Kits offer a proprietary calcium assay formulation that requires neither a wash step nor aRead more
| Catalog Number | Quantity |
|---|---|
| F36206 | 10 Microplates |
Catalog number F36206
Price (EUR)
479,00
Each
Quantity:
10 Microplates
Price (EUR)
479,00
Each
Fluo-4 NW (No-Wash) Calcium Assay Kits offer a proprietary calcium assay formulation that requires neither a wash step nor a quencher dye. The fluo-4 NW assay achieves larger increases in fluorescence intensity than standard fluo-3 and fluo-4 assays with a wash step. Eliminating the wash step results in lower variability and higher Z´ values than the standard fluo-4 assay, while providing an easier and faster assay as well.
The fluo-4NW indicator is nonfluorescent and stable in pH 7–7.5 buffer for several hours, so spontaneous conversion to the Ca2+-sensitive form is not a significant source of background fluorescence. Contributions to baseline fluorescence by the growth medium (e.g., esterase activity, proteins interacting with receptors of interest, or phenol red) are eliminated by removing the medium prior to adding the indicator dye to the wells.
Another source of potential fluorescence outside the cells is extrusion of the indicator out of the cell by organic anion transporters. Probenecid is commonly used to inhibit this transport and reduce the baseline signal. We have synthesized a proprietary water-soluble probenecid, which is supplied with the Fluo-4 NW Calcium Assay Kits. This form of probenecid has the advantages of being easy to dissolve in buffer and safer to use than the free acid, which requires caustic 1 M NaOH to dissolve. TheFluo-4 NW Calcium Assay Kits are designed for microplates and HTS, and the assay can be performed on adherent as well as nonadherent cells.
Fluo-4 AM is a fluorescent Ca+2 indicator that is widely used for in-cell measurement of agonist-stimulated and antagonist inhibited calcium signaling in high-throughput screening (HTS) applications. Its visible wavelength excitation (compatible with argon-ion laser sources), high sensitivity, and large fluorescence increase upon binding Ca2+ has made it the indicator of choice for characterizing G-protein–coupled receptor (GPCR) pharmacology and function. These properties have made fluo-4 AM attractive not only for microplate screening applications but for microscopy and flow cytometry as well.
The fluo-4NW indicator is nonfluorescent and stable in pH 7–7.5 buffer for several hours, so spontaneous conversion to the Ca2+-sensitive form is not a significant source of background fluorescence. Contributions to baseline fluorescence by the growth medium (e.g., esterase activity, proteins interacting with receptors of interest, or phenol red) are eliminated by removing the medium prior to adding the indicator dye to the wells.
Another source of potential fluorescence outside the cells is extrusion of the indicator out of the cell by organic anion transporters. Probenecid is commonly used to inhibit this transport and reduce the baseline signal. We have synthesized a proprietary water-soluble probenecid, which is supplied with the Fluo-4 NW Calcium Assay Kits. This form of probenecid has the advantages of being easy to dissolve in buffer and safer to use than the free acid, which requires caustic 1 M NaOH to dissolve. TheFluo-4 NW Calcium Assay Kits are designed for microplates and HTS, and the assay can be performed on adherent as well as nonadherent cells.
Fluo-4 AM is a fluorescent Ca+2 indicator that is widely used for in-cell measurement of agonist-stimulated and antagonist inhibited calcium signaling in high-throughput screening (HTS) applications. Its visible wavelength excitation (compatible with argon-ion laser sources), high sensitivity, and large fluorescence increase upon binding Ca2+ has made it the indicator of choice for characterizing G-protein–coupled receptor (GPCR) pharmacology and function. These properties have made fluo-4 AM attractive not only for microplate screening applications but for microscopy and flow cytometry as well.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
Dye TypeFluorescent Dye-Based
Quantity10 Microplates
Shipping ConditionRoom Temperature
For Use With (Application)Calcium Assay
For Use With (Equipment)Microplate Reader
Product TypeStain
Unit SizeEach
Contents & Storage
Store in freezer -5°C to -30°C and protect from light.
Citations & References (21)
Citations & References
Abstract
Cell death and autophagy under oxidative stress: roles of poly(ADP-Ribose) polymerases and Ca(2+).
Journal:Mol Cell Biol
PubMed ID:22751932
On the cellular level, oxidative stress may cause various responses, including autophagy and cell death. All of these outcomes involve disturbed Ca(2+) signaling. Here we show that the nuclear enzymes poly(ADP-ribose) polymerase 1 (PARP1) and PARP2 control cytosolic Ca(2+) shifts from extracellular and intracellular sources associated with autophagy or cell
The Nef protein of human immunodeficiency virus is a broad-spectrum modulator of chemokine receptor cell surface levels that acts independently of classical motifs for receptor endocytosis and Galphai signaling.
Journal:Mol Biol Cell
PubMed ID:16775006
'Chemokine receptors (CKRs) are important physiological mediators of immune defense, inflammatory responses, and angiogenesis, and they have also been implicated in a number of viral disease processes. Here, we report that the Nef protein of human immunodeficiency virus (HIV) reduces cell surface levels of eight different members of the CC-
Development and validation of a cell-based high-throughput screening assay for TRPM2 channel modulators.
Journal:J Biomol Screen
PubMed ID:18057180
'TRPM2 is a member of the transient receptor potential melastatin (TRPM)-related ion channel family. The activation of TRPM2 induced by oxidative/nitrosative stress leads to an increase in intracellular free Ca(2+). Although further progress in understanding TRPM2''s role in cell and organism physiology would be facilitated by isolation of compounds able
Fc receptor-like 5 inhibits B cell activation via SHP-1 tyrosine phosphatase recruitment.
Journal:Proc Natl Acad Sci U S A
PubMed ID:17522256
'The Fc receptor-like protein 5 (FCRL5) on B cells has both an immunoreceptor tyrosine-based activation motif (ITAM)-like sequence and two consensus immunoreceptor tyrosine-based inhibitory motifs (ITIM) in its cytoplasmic region. To evaluate its signaling potential, we expressed constructs for chimeric molecules composed of the cytoplasmic region of FCRL5 and the
Pertussis toxin signals through the TCR to initiate cross-desensitization of the chemokine receptor CXCR4.
Journal:J Immunol
PubMed ID:19380820
Pertussis toxin (PTx) has been shown to exert a variety of effects on immune cells independent of its ability to ADP-ribosylate G proteins. Of these effects, the binding subunit of PTx (PTxB) has been shown to block signaling via the chemokine receptor CCR5, but the mechanism involved in this process