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

Catalog No. C3015
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C3015 10 x 50 μg
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Catalog No. C3015 Supplier Invitrogen™ Supplier No. C3015
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Description

Labeled calcium indicators are molecules that exhibit an increase in fluorescence upon binding Ca2+

  • These indicators 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
  • Fluorescence signal from these cells is generally measured using fluorescence microscopy, fluorescence microplate assays, or flow cytometry
  • Exhibit fluorescence increase upon binding Ca2+ with little shift in wavelength
  • These probes are excited by visible light, and because energy required for excitation is low, 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
  • Label (Ex/Em): Calcium Orange (549/576nm)
  • Fluorescence intensity increase upon binding Ca2+ : approximately 3 fold

Calcium Detection, Cell Analysis, Cell Viability, Proliferation and Function, Ionic Homeostasis and Signaling

Order Info

Shipping Condition: Room temperature

Specifications

Content And Storage Store in freezer -5°C to -30°C and protect from light.
Detection Method Fluorescence
For Use With (Application) Cell Viability and Proliferation
For Use With (Equipment) Fluorescence Microscope
Product Type Calcium Indicator
Dye Type Fluorescent Dye-Based
Product Line Calcium Orange
Quantity 10 x 50 μg
Shipping Condition Room Temperature

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).
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.
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.

For Research Use Only. Not for use in diagnostic procedures.