Fluo-4 Direct™ Calcium Assay Kit
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Citations & References (8)
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
F104721 x 100 mL
F1047110 x 10 mL
F104731 x 1 L
Catalog number F10472
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4,951.00
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Ends: 31-Dec-2025
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Quantity:
1 x 100 mL
Price (CNY)
4,951.00
飞享价
Ends: 31-Dec-2025
6,493.00
Save 1,542.00 (24%)
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
Quantity1 x 100 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
  • 100 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 (8)

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
Transient receptor potential vanilloid-1 (TRPV1) is a mediator of lung toxicity for coal fly ash particulate material.
Authors:Deering-Rice CE, Johansen ME, Roberts JK, Thomas KC, Romero EG, Lee J, Yost GS, Veranth JM, Reilly CA,
Journal:Mol Pharmacol
PubMed ID:22155782
'Environmental particulate matter (PM) pollutants adversely affect human health, but the molecular basis is poorly understood. The ion channel transient receptor potential vanilloid-1 (TRPV1) has been implicated as a sensor for environmental PM and a mediator of adverse events in the respiratory tract. The objectives of this study were to ... 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
Screening ß-arrestin recruitment for the identification of natural ligands for orphan G-protein-coupled receptors.
Authors:Southern C, Cook JM, Neetoo-Isseljee Z, Taylor DL, Kettleborough CA, Merritt A, Bassoni DL, Raab WJ, Quinn E, Wehrman TS, Davenport AP, Brown AJ, Green A, Wigglesworth MJ, Rees S,
Journal:J Biomol Screen
PubMed ID:23396314
A variety of G-protein-coupled receptor (GPCR) screening technologies have successfully partnered a number of GPCRs with their cognate ligands. GPCR-mediated ß-arrestin recruitment is now recognized as a distinct intracellular signaling pathway, and ligand-receptor interactions may show a bias toward ß-arrestin over classical GPCR signaling pathways. We hypothesized that the failure ... More
A novel in vitro model system for smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells.
Authors:Guo X, Stice SL, Boyd NL, Chen SY,
Journal:Am J Physiol Cell Physiol
PubMed ID:23220114
The objective of this study was to develop a novel in vitro model for smooth muscle cell (SMC) differentiation from human embryonic stem cell-derived mesenchymal cells (hES-MCs). We found that hES-MCs were differentiated to SMCs by transforming growth factor-ß (TGF-ß) in a dose- and time-dependent manner as demonstrated by the ... More
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