Fluo-5N, AM, cell permeant - Special Packaging
Fluo-5N, AM, cell permeant - Special Packaging
Invitrogen™

Fluo-5N, AM, cell permeant - Special Packaging

Labeled calcium indicators are molecules that exhibit an increase in fluorescence upon binding Ca2+. Fluo-5F, fluo-5N, and fluo-4ff are analogsRead more
Catalog NumberQuantity
F1420410 x 50 μg
Catalog number F14204
Price (JPY)
88,200
Each
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Quantity:
10 x 50 μg
Labeled calcium indicators are molecules that exhibit an increase in fluorescence upon binding Ca2+. Fluo-5F, fluo-5N, and fluo-4ff are analogs of fluo-4 with lower Ca2+-binding affinity, making them suitable for detecting intracellular calcium levels in the 1 μM to 1 mM range that would saturate the response of fluo-3 and fluo-4. Cells may be loaded with the AM ester forms of these calcium indicators by adding the dissolved indicator directly to dishes containing cultured cells. These indicators are compatible with excitation at 488 nm by argon-ion laser sources, making them useful for confocal microscopy, flow cytometry, and microplate screening applications.

Learn more about ion indicators including calcium, potassium, pH, and membrane potential indicators ›

Calcium Indicator (AM Ester) Specifications:
• Label (Ex/Em of Ca2+–bound form): Fluo-5N (494/516 nm)
• Fluorescence intensity increase upon binding Ca2+: >100 fold
• Kd for Ca2+ in buffer: ∼90 μM
• Exhibit fluorescence increase upon binding Ca2+ with little shift in wavelength


Using TPEN to Control Heavy Metal Cations
In addition, BAPTA-based indicators such as these bind various heavy metal cations (e.g., Mn2+, Zn2+, Pb2+) with substantially higher affinity than Ca2+. Perturbations to calcium measurements caused by presence of these ions can be controlled using the heavy metal-selective chelator TPEN.

More Choices for Fluorescent Calcium Indicators
We offer a large selection of Molecular Probes™ calcium indicators for use in various experimental scenarios. For more information, review Fluorescent Ca2+ Indicators Excited with Visible Light—Section 19.3 in the Molecular Probes™ Handbook.

For UV-excitable Ca2+ indicators, protein-based Ca2+ indicators, conjugates of Ca2+ indicators, and for fluorescence-based indicators of other metal ions (i.e., Mg2+, Zn2+) review Indicators for Ca2+, Mg2+, Zn2+ and Other Metal Ions—Chapter 19 in the Molecular Probes™ Handbook.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
Dye TypeFluorescent Dye-Based
Quantity10 x 50 μg
Shipping ConditionRoom Temperature
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 TypeCalcium Indicator
Unit SizeEach
Contents & Storage
Store in freezer -5°C to -30°C 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 (15)

Citations & References
Abstract
The use of the indicator fluo-5N to measure sarcoplasmic reticulum calcium in single muscle fibres of the cane toad.
Authors:Kabbara AA, Allen DG
Journal:J Physiol
PubMed ID:11432994
'1. Single fibres from the lumbrical muscles of the cane toad (Bufo marinus) were incubated in fluo-5N AM for 2 h at 35 degrees C in order to load the indicator into the sarcoplasmic reticulum. Fluo-5N is a low-affinity calcium indicator (K(Ca) 90 microM). Successful sarcoplasmic reticulum (SR) loading was ... More
Cardiac alternans do not rely on diastolic sarcoplasmic reticulum calcium content fluctuations.
Authors:Picht E, DeSantiago J, Blatter LA, Bers DM,
Journal:Circ Res
PubMed ID:16946134
'Cardiac alternans are thought to be a precursor to life-threatening arrhythmias. Previous studies suggested that alterations in sarcoplasmic reticulum (SR) Ca2+ content are either causative or not associated with myocyte Ca2+ alternans. However, those studies used indirect measures of SR Ca2+. Here we used direct continuous measurement of intra-SR free ... More
Kinesin dependent, rapid, bi-directional transport of ER sub-compartment in dendrites of hippocampal neurons.
Authors:Bannai H, Inoue T, Nakayama T, Hattori M, Mikoshiba K
Journal:J Cell Sci
PubMed ID:14676272
'Although spatially restricted Ca2+ release from the endoplasmic reticulum (ER) through intracellular Ca2+ channels plays important roles in various neuronal activities, the accurate distribution and dynamics of ER in the dendrite of living neurons still remain unknown. To elucidate these, we expressed fluorescent protein-tagged ER proteins in cultured mouse hippocampal ... More
Ca2+ blinks: rapid nanoscopic store calcium signaling.
Authors:Brochet DX, Yang D, Di Maio A, Lederer WJ, Franzini-Armstrong C, Cheng H
Journal:Proc Natl Acad Sci U S A
PubMed ID:15710901
'Luminal Ca(2+) in the endoplasmic and sarcoplasmic reticulum (ER/SR) plays an important role in regulating vital biological processes, including store-operated capacitative Ca(2+) entry, Ca(2+)-induced Ca(2+) release, and ER/SR stress-mediated cell death. We report rapid and substantial decreases in luminal [Ca(2+)], called "Ca(2+) blinks," within nanometer-sized stores (the junctional cisternae of ... More
Sulfhydryl oxidation overrides Mg(2+) inhibition of calcium-induced calcium release in skeletal muscle triads.
Authors:Donoso P, Aracena P, Hidalgo C
Journal:Biophys J
PubMed ID:10866954
'We studied the effect of oxidation of sulfhydryl (SH) residues on the inhibition by Mg(2+) of calcium-induced calcium release (CICR) in triad-enriched sarcoplasmic reticulum vesicles isolated from rabbit skeletal muscle. Vesicles were either passively or actively loaded with calcium before eliciting CICR by dilution at pCa 4.6-4.4 in the presence ... More