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| Product Name | Indo-1 AM Calcium Indicator (Cell Permeant) |
| Catalog No. | RESK-0565 |
| Description | Indo-1 is one of the most widely used ratiometric fluorescent calcium indicators in cell biology, alongside Fura-2. It binds Ca2+ specifically and fluoresces; upon Ca2+ binding its emission maximum shifts from 475 nm (Ca2+-free) toward 400 nm (Ca2+-saturated). The probe is excited at a single wavelength in the 351-364 nm range of an argon-ion laser, and the fluorescence signals of the Ca2+-bound and Ca2+-free forms are detected at 400 nm and 475 nm, respectively; the ratio of the two intensities is used to determine calcium concentration. Because Indo-1 is a highly polar acidic molecule that cannot cross the cell membrane, its anionic groups are masked with acetoxymethyl (AM) esters to yield Indo-1 AM. This modification increases lipophilicity and abolishes the negative charge, greatly enhancing cellular uptake. Inside the cell, Indo-1 AM is hydrolyzed by intracellular esterases to Indo-1, which reversibly binds free cytosolic Ca2+. |
| Intended Use | Ratiometric measurement of intracellular free Ca2+ concentration in live cells, particularly by flow cytometry, for studying changes in the intracellular environment. |
| Principle / Technology | Ratiometric fluorescent calcium indicator. The cell-permeant acetoxymethyl (AM) ester enters live cells and is hydrolyzed by intracellular esterases to the Ca2+-sensitive Indo-1. Ca2+ binding shifts the emission maximum from 475 nm to 400 nm, and the 400/475 nm emission ratio reflects cytosolic Ca2+ concentration. |
| Detection Method | Fluorescence-based ratiometric detection; single-wavelength excitation at 351-364 nm (375 nm excitation is also feasible) with dual emission readout at 400 nm and 475 nm; well suited to flow cytometry. |
| Sample Type | Live cells; no fixation is required before probe loading. |
| Performance Range / Specifications | Emission maximum shifts from 475 nm (Ca2+-free) to approximately 400 nm upon Ca2+ saturation, enabling ratiometric quantification of Ca2+. |
| Sensitivity / LOD | Highly sensitive; effectively detects changes in Ca2+ concentration to support prediction and study of intracellular environmental changes. |
| Specificity | High specificity for Ca2+, with minimal interference from other ions that may be present. |
| Reaction Conditions / Protocol | Load live cells with Indo-1 AM; intracellular esterases hydrolyze the probe to active Indo-1, which reversibly binds cytosolic free Ca2+. Excite at a single wavelength in the 351-364 nm range (375 nm is also usable) and record emission at 400 nm and 475 nm for ratio calculation. Cells do not need to be fixed prior to staining. |
| Components / Formulation | Indo-1 acetoxymethyl (AM) ester, cell permeant. |
| Storage Conditions | Store at -20 °C, protected from light and kept dry. |
| Shelf Life | 1 year when stored as recommended. |
| Package Specifications | 50 μg; 5 x 50 μg |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | Stable for 1 year when stored at -20 °C, protected from light and dry. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Compatible with cells transfected with EGFP- or EYFP-expressing plasmids, which do not interfere with detection; detectable on flow cytometers equipped with a 351-364 nm argon-ion laser line. |
| Application Notes / Precautions | The fluorescence color changes from blue to violet upon Ca2+ binding. Use with live, unfixed cells. Protect the probe from light. |
For research use only, not for clinical use.
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