Fluo-4 Direct™ Calcium Assay Kit
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Citations & References (13)
Invitrogen™

Fluo-4 Direct™ Calcium Assay Kit

The Fluo-4 Direct™ Calcium Assay Kit was formulated to provide a homogeneous fluorescent calcium assay that: 1. Easily loads intoRead more
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Catalog NumberQuantity
F1047110 x 10 mL
F104721 x 100 mL
F104731 x 1 L
Catalog number F10471
Price (BRL)
5.453,65
Each
Add to cart
Quantity:
10 x 10 mL
Price (BRL)
5.453,65
Each
Add to cart
The Fluo-4 Direct™ Calcium Assay Kit was formulated to provide a homogeneous fluorescent calcium assay that:
1. Easily loads into cells
2. Achieves a large assay window and
3. Suppresses background fluorescence generated from the calcium indicator in complete media with little-to-no impact on the specific cellular fluorescence

This advanced formulation allows the assay to be run in a simple “addition only” format in the presence of serum-containing media. Fluo-4 Direct™ is the third addition to the Fluo-4 family of calcium detection reagents. Fluo-4 AM and Fluo-4 NW both require media removal before assay detection, but Fluo-4 NW adds convenience by including the PowerLoad™ reagent for ease in cell loading. The Fluo-4 Direct™ Calcium Assay Kit is similar to Fluo-4 NW in that it is formulated with PowerLoad™ for ease in cell loading, but unique in that it can be used in the presence of complete culture media and will efficiently suppress background fluorescence without sacrificing the specific cellular fluorescence generated in the assay.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
Dye TypeFluorescent Dye-Based
FormPowder
ModificationChemical
Quantity10 x 10 mL
Shipping ConditionRoom Temperature
Sub Cellular LocalizationNucleus, Organelles, Cytoplasm
ColorGreen
EmissionVisible
For Use With (Application)Calcium Assay
For Use With (Equipment)Confocal Microscope, Fluorescence Microscope, High Content Analysis Instrument, HTS Reader, Microplate Reader, Fluorescent Imager
Product LineMolecular Probes
Product TypeCalcium Assay Kit
Unit SizeEach
Contents & Storage
  • 10 × 10 ml Fluo-4 Direct™ Calcium Assay Reagent (Component A)
  • 2 × 77 mg Probenecid (Component B)
  • 200 ml Fluo-4 Direct™ Calcium Assay Buffer (Component C)
  • Store at ≤-20°C. Dessicate and protect from light.

Frequently asked questions (FAQs)

I am doing calcium flux imaging with your Fura-2 calibration kit, but am seeing a large variability in ratio in different places around the slide. I am correcting for uniform illumination, using the product as directed, and sealing the coverslip with nail polish.

The nail polish may be the problem. The Kd value (calcium sensitivity) changes depending upon the dye's environment. Nail polish has solvents that can leech under the coverslip and cause variability. We recommend either going without a sealing or sealing with melted paraffin painted on the coverslip edges with a cotton-tipped applicator (paraffin is hydrophobic and has no solvents).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I need to label cells with Fluo-4, AM, for a calcium flux assay. How long after labeling will the dye be retained?

After loading dye into the cells, intracellular esterases remove the 'AM' moiety from the dye. When the 'AM' group is removed, the dye is able to bind calcium and fluoresce. Since the dye is not covalently bound to any cellular components, it may be actively effluxed from the cell. The rate of efflux is dependent upon the inherent properties of the cell, culture conditions and other factors. The dye may be retained for hours, days or even weeks or lost in a matter of minutes. The use of Probenecid (Cat. No. P36400) limits loss by active efflux.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Citations & References (13)

Citations & References
Abstract
High-resolution imaging of the immunological synapse and T-cell receptor microclustering through microfabricated substrates.
Authors:Biggs MJ, Milone MC, Santos LC, Gondarenko A, Wind SJ,
Journal:J R Soc Interface
PubMed ID:21490003
'T-cell activation via antigen presentation is associated with the formation of a macromolecular membrane assembly termed the immunological synapse (IS). The genesis of the IS and the onset of juxtacrine signalling is characterized by the formation of cell membrane microclusters and the organization of such into segregated microdomains. A central ... More
Regulation of cytosolic phospholipase A2 (cPLA2alpha) and its association with cell proliferation in human lens epithelial cells.
Authors:Wang Y, Xing KY, Lou MF,
Journal:Invest Ophthalmol Vis Sci
PubMed ID:21896865
'To investigate the molecular mechanism for cytosolic phospholipase A2 (cPLA(2)a) regulation and its association to platelet-derived growth factor (PDGF)-induced cell proliferation. cPLA(2)a was examined using human lens epithelial (HLE) B3 cells. Reactive oxygen species (ROS) generation induced by PDGF was analyzed by luminescence assay. Cell proliferation was measured by cell ... More
20-Hydroxyeicosatetraenoic acid (20-HETE) is a novel activator of transient receptor potential vanilloid 1 (TRPV1) channel.
Authors:Wen H, Östman J, Bubb KJ, Panayiotou C, Priestley JV, Baker MD, Ahluwalia A,
Journal:J Biol Chem
PubMed ID:22389490
'TRPV1 is a member of the transient receptor potential ion channel family and is gated by capsaicin, the pungent component of chili pepper. It is expressed predominantly in small diameter peripheral nerve fibers and is activated by noxious temperatures >42 °C. 20-Hydroxyeicosatetraenoic acid (20-HETE) is a cytochrome P-450 4A/4F-derived metabolite ... More
Gq/5-HT2c receptor signals activate a local GABAergic inhibitory feedback circuit to modulate serotonergic firing and anxiety in mice.
Authors:Spoida K, Masseck OA, Deneris ES, Herlitze S,
Journal:
PubMed ID:24733892
Serotonin 2c receptors (5-HT2c-Rs) are drug targets for certain mental disorders, including schizophrenia, depression, and anxiety. 5-HT2c-Rs are expressed throughout the brain, making it difficult to link behavioral changes to circuit specific receptor expression. Various 5-HT-Rs, including 5-HT2c-Rs, are found in the dorsal raphe nucleus (DRN); however, the function of ... More
Small molecule inhibition of PAX3-FOXO1 through AKT activation suppresses malignant phenotypes of alveolar rhabdomyosarcoma.
Authors:Jothi M, Mal M, Keller C, Mal AK,
Journal:
PubMed ID:24107448
Alveolar rhabdomyosarcoma comprises a rare highly malignant tumor presumed to be associated with skeletal muscle lineage in children. The hallmark of the majority of alveolar rhabdomyosarcoma is a chromosomal translocation that generates the PAX3-FOXO1 fusion protein, which is an oncogenic transcription factor responsible for the development of the malignant phenotype ... More
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