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Invitrogen™ Rhod-2, AM, cell permeant

Catalog No. R1244
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20 x 50 μg
1 mg
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Catalog No. Quantity
R1244 1 mg
R1245MP 20 x 50 μg
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Catalog No. R1244 Supplier Invitrogen™ Supplier No. R1244

Description

Cells may be loaded with AM ester forms of these calcium indicators by adding dissolved indicator directly to dishes containing cultured cells

Labeled calcium indicators are molecules that exhibit increase in fluorescence upon binding Ca2+. They have uses in many calcium signaling investigations, including measuring Ca2+ in cells and tissues that have high levels of autofluorescence and also for detecting Ca2+ release generated by photoreceptors and photoactivatable chelators.

  • Fluorescence signal from these cells is generally measured using fluorescence microscopy
  • Label (Ex/Em of Ca2+-bound form): Rhod-2 (552/581nm)
  • Fluorescence intensity increase upon binding Ca2+: >100 fold
  • Kd for Ca2+ in the absence of Mg2+, in buffer: approximately 570nM
  • Exhibit fluorescence increase upon binding Ca2+ with little shift in wavelength
  • 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

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 Stain
Dye Type Fluorescent Dye-Based
Quantity 1 mg
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).
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.

What are the excitation/emission maxima for Rhod-2, AM, cell permeant (Cat. No. R1244, R1245MP)?

Rhod-2, AM, cell permeant (Cat. No. R1244, R1245MP) exhibits >100-fold increase in fluorescence intensity upon binding Ca2+. The excitation/emission maxima for Rhod-2, AM, cell permeant when bound to Ca2+ are 552/581 nm.

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