Fluo-4 Direct™ Calcium Assay Kit

Citations & References (8)

Invitrogen™

Fluo-4 Direct™ Calcium Assay Kit

Name: Fluo-4 Direct™ Calcium Assay Kit
SKU: F10473

The Fluo-4 Direct™ Calcium Assay Kit was formulated to provide a homogeneous fluorescent calcium assay that: 1. Easily loads intoRead more

Change view

Catalog Number	Quantity
F10473	1 x 1 L
F10471	10 x 10 mL
F10472	1 x 100 mL

3 Options

Catalog number F10473

Price (MXN)

Quantity:

1 x 1 L

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 1 L

Shipping ConditionRoom Temperature

Sub Cellular LocalizationNucleus, Organelles, Cytoplasm

ColorGreen

EmissionVisible

For Use With (Application)Cell Viability and Proliferation

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

1 L Fluo-4 Direct™ Calcium Assay Reagent (Component A)
1.54 g Probenecid (Component B)
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 K_d 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

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

Selective Impairment of P2Y Signaling by Prostaglandin E2 in Macrophages: Implications for Ca2+-Dependent Responses.

Authors:Través PG, Pimentel-Santillana M, Carrasquero LM, Pérez-Sen R, Delicado EG, Luque A, Izquierdo M, Martín-Sanz P, Miras-Portugal MT, Boscá L,

Journal:J Immunol

PubMed ID:23479225

Extracellular nucleotides have been recognized as important modulators of inflammation via their action on specific pyrimidine receptors (P2). This regulation coexists with the temporal framework of proinflammatory and proresolution mediators released by the cells involved in the inflammatory response, including macrophages. Under proinflammatory conditions, the expression of cyclooxygenase-2 leads to ... More

Surfactant protein A integrates activation signal strength to differentially modulate T cell proliferation.

Authors:Mukherjee S, Giamberardino C, Thomas J, Evans K, Goto H, Ledford JG, Hsia B, Pastva AM, Wright JR,

Journal:J Immunol

PubMed ID:22219327

Pulmonary surfactant lipoproteins lower the surface tension at the alveolar-airway interface of the lung and participate in host defense. Previous studies reported that surfactant protein A (SP-A) inhibits lymphocyte proliferation. We hypothesized that SP-A-mediated modulation of T cell activation depends upon the strength, duration, and type of lymphocyte activating signals. ... More

HIV-1 Tat protein inhibits neurosecretion by binding to phosphatidylinositol 4,5-bisphosphate.

Authors:Tryoen-Tóth P, Chasserot-Golaz S, Tu A, Gherib P, Bader MF, Beaumelle B, Vitale N,

Journal:J Cell Sci

PubMed ID:23178941

HIV-1 transcriptional activator (Tat) enables viral transcription and is also actively released by infected cells. Extracellular Tat can enter uninfected cells and affect some cellular functions. Here, we examine the effects of Tat protein on the secretory activity of neuroendocrine cells. When added to the culture medium of chromaffin and ... More

View all Fluo-4 Direct™ Calcium Assay Kit, 1 x 1 L citations