Contents: Indo-1 AM Calcium Sensor Dye Catalog Number: 65-0856 Formulation: Lyophilized yellow solid. Purity > 96% as determined by HPLC. Storage Conditions: Store at -20°C. Protect from light and moisture.
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65-0856-39
1 mg
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
Indo-1 AM Calcium Sensor Dye is a membrane-permeable dye used for determining changes in calcium concentrations in the cell using fluorescence microscopy, flow cytometry, fluorescence spectroscopy and fluorescence microplate readers. Once Indo-1 enters the cell, esterases cleave the AM group yielding a membrane-impermeable dye. Unbound Indo-1 has a peak emission at 485 nm. Upon binding calcium, the peak emission shifts down to 410 nm.
In flow cytometry this can be measured over time and can be represented as a ratio of the two emission wavelengths. To optimize the ratio between the two emissions, unbound Indo-1 fluorescence should be collected using a filter above 485nm (525 nm is a good option) while bound Indo-1 fluorescence should be collected using a filter below 400 nm. Because the emission profile of Indo-1 is broad, multicolor flow analysis using eFluor TM 450, eFluorTM605NC or eFluorTM 650NC is not possible, however fluorochrome conjugated antibodies utilizing the 488nm or 633 nm are quite compatible with Indo-1.
Indo-1 AM should be reconstituted in high-quality, freshly opened DMSO. Recommended concentration: 1 mM. Once reconstituted, it should be protected from light and stored at -20°C; avoid freeze-thaws.
Applications Reported
For research use only, not for diagnostic or therapeutic use. Indo-1 AM has been reported for use in flow cytometric analysis.
Applications Tested
It is recommended that Indo-1 AM be used at concentrations between 0.1 - 1 µM in flow cytometric applications. It is highly recommended that the concentration and labeling conditions be carefully determined by each investigator for optimal performance in the assay of interest.
References
Jennings LK, Dockter ME, Wall CD, Fox CF, Kennedy DM. Calcium mobilization in human platelets using indo-1 and flow cytometry. Blood. 1989 Dec;74(8):2674-80.
