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Invitrogen™ Rhod-2, AM, cell permeant
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 an 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. 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.
Calcium Indicator (AM Ester) Specifications:
- Label (Ex/Em of Ca2+–bound form): Rhod-2 (552/581 nm)
- Fluorescence intensity increase upon binding Ca2+: >100 fold
- Kd for Ca2+ in the absence of Mg2+, in buffer: ∼570 nM
- 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 (Cat. T1210).
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 for human or animal therapeutic or diagnostic use.
Order Info
Shipping Condition: Room temperature
Specifications
Specifications
| Content And Storage | Store in freezer -5°C to -30°C and protect from light. |
| Detection Method | Fluorescence |
| For Use With (Application) | Calcium Indicator |
| For Use With (Equipment) | Fluorescence Microscope |
| Product Type | Stain |
| Dye Type | Rhod-2 |
| Quantity | 1 mg |
| Shipping Condition | Room Temperature |
| Excitation Wavelength | 552 nm |
| Emission Wavelength | 581 nm |
Frequently Asked Questions (FAQs)
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).
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.
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.