Calcium Orange™, AM, cell permeant - Special Packaging
Calcium Orange™, AM, cell permeant - Special Packaging
Invitrogen™

Calcium Orange™, AM, cell permeant - Special Packaging

Labeled calcium indicators are molecules that exhibit an increase in fluorescence upon binding Ca2+. They have uses in many calciumLeia mais
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Número do catálogoQuantity
C3015
conhecido também como C-3015
10 x 50 μg
Número do catálogo C3015
conhecido também como C-3015
Preço (BRL)
-
Quantity:
10 x 50 μg
Labeled calcium indicators are molecules that exhibit an increase in fluorescence upon binding Ca2+. They have uses in many calcium signaling investigations, including measuring intracellular Ca2+, following Ca2+ influx and release, and multiphoton excitation imaging of Ca2+ in living tissues. Cells may be loaded with the AM ester forms of these calcium indicators by adding the dissolved indicator directly to dishes containing cultured cells. The fluorescence signal from these cells is generally measured using fluorescence microscopy, fluorescence microplate assays, or flow cytometry.

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

Calcium Indicator (AM Ester) Specifications:
• Label (Ex/Em): Calcium Orange™ (549/576 nm)
• Fluorescence intensity increase upon binding Ca2+ : ∼3 fold
• Exhibit fluorescence increase upon binding Ca2+ with little shift in wavelength


Spectral Characteristics of Molecular Probes™ Calcium Indicators
These probes are excited by visible light, and because the energy required for excitation is low, the potential for cellular photodamage is reduced. Commonly used laser-based instruments (i.e., confocal laser scanning microscopes) are able to efficiently excite these indicators, and their emissions are in regions of the spectrum where cellular autofluorescence and scattering backgrounds are often less of a problem.

More Choices for Fluorescent Calcium Indicators
We offer a large selection of Molecular Probes™ calcium indicators for use in various experimental scenarios, for example dextran versions for reduced leakage and compartmentalization and BAPTA conjugates for detecting high-amplitude calcium transients. 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 intended for animal or human therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.
Especificações
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)Fluorescence Microscope
Product LineCalcium Orange
Product TypeCalcium Indicator
Unit SizeEach
Conteúdo e armazenamento
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.

When using Calcium Crimson, AM, as an indicator of intracellular calcium flux, I am not getting a good degree of change. The cells also have GFP. What can I do?

This is a known drawback of Calcium Crimson, AM (as well as Calcium Orange, AM and Fura Red, AM, which are also in the same emission range). You can try increasing the concentration and washing out of any unlabeled dye from the media, to try to get better signal-to-background. If that fails, we recommend using Rhod-3 AM instead, which has a much better change in signal in that wavelength.

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.

Citações e referências (28)

Citações e referências
Abstract
AM-loading of fluorescent Ca2+ indicators into intact single fibers of frog muscle.
Authors:Zhao M, Hollingworth S, Baylor SM
Journal:Biophys J
PubMed ID:9168048
'The AM loading of a number of different fluorescent Ca2+ indicators was compared in intact single fibers of frog muscle. Among the 13 indicators studied, loading rates (the average increase in the fiber concentration of indicator per first 60 min of loading) varied approximately 100-fold, from approximately 3 microM/h to ... More
Intracellular Ca2+ dynamics during spontaneous and evoked activity of leech heart interneurons: low-threshold Ca currents and graded synaptic transmission.
Authors:Ivanov AI, Calabrese RL
Journal:J Neurosci
PubMed ID:10864951
'In oscillatory neuronal networks that pace rhythmic behavior, Ca(2+) entry through voltage-gated Ca channels often supports bursting activity and mediates graded transmitter release. We monitored simultaneously membrane potential and/or ionic currents and changes of Ca fluorescence (using the fluorescence indicator Ca Orange) in spontaneously active and experimentally manipulated oscillator heart ... More
The role of Ca(2+) in stimulated bioluminescence of the dinoflagellate Lingulodinium polyedrum.
Authors:von Dassow P, Latz MI
Journal:J Exp Biol
PubMed ID:12200401
'Many marine dinoflagellates emit bright discrete flashes of light nearly instantaneously in response to either laminar or turbulent flows as well as to direct mechanical stimulation. The flash involves a unique pH-dependent luciferase and a proton-mediated action potential across the vacuole membrane. The mechanotransduction process initiating this action potential is ... More
Ryanodine receptor mutations associated with stress-induced ventricular tachycardia mediate increased calcium release in stimulated cardiomyocytes.
Authors:George CH, Higgs GV, Lai FA
Journal:Circ Res
PubMed ID:12919952
'Ca2+ release from the sarcoplasmic reticulum mediated by the cardiac ryanodine receptor (RyR2) is a fundamental event in cardiac muscle contraction. RyR2 mutations suggested to cause defective Ca2+ channel function have recently been identified in catecholaminergic polymorphic ventricular tachycardia (CPVT) and arrhythmogenic right ventricular dysplasia (ARVD) affected individuals. We report ... More
Cell-permeant Ca2+ chelators reduce early excitotoxic and ischemic neuronal injury in vitro and in vivo.
Authors:Tymianski M, Wallace MC, Spigelman I, Uno M, Carlen PL, Tator CH, Charlton MP
Journal:Neuron
PubMed ID:8102532
'We report the characterization of the first successful treatment of neuronal ischemic injury in vivo by cell-permeant Ca2+ chelators. The chelators attenuated glutamate-induced intracellular Ca2+ increases and neurotoxicity in neuronal explant cultures. When infused intravenously in rats, permeant fluorescent BAPTA analogs accumulated in neurons in several brain regions. BAPTA-AM, infused ... More