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Calcein Deep Red™ AM ester
Calcein AM is one the most popular fluorescent probes used for labeling and monitoring cellular functions of live cells. However, the single color of Calcein AM makes it impossible to use this valuable reagent in the multicolor applications. For example, it is impossible to use Calcein AM in combination of GFP-tranfacted cells due to the same color to GFP. To address this color limitation of Calcein AM, we have developed Calcein Orange™, Calcein Red™ and Calcein Deep Red™. These new Calcein AM ester enables the multicolor labeling and functional analysis of live cells in combination with Calcein AM. Non-fluorescent Calcein Deep Red™ AM ester can easily get into live cells and hydrolyzes to generate strongly fluorescent Calcein Deep Red™ (Cat#: 21902) dye. Calcein Deep Red™ dye can be monitored with the common Cy5 filter set.
Flow Cytometry Analysis of Jurkat cells stained with Calcein Deep Red™ AM ester (Cat#22011). Jurkat cells were washed once with HH buffer and stained with 2 uM Calcein Deep Red™ AM ester (Cat#22011) in HH with 0.02% PF-127(Cat#20053) and 1mM PBC (Cat# 20061) for 30 minutes at 37C incubator. Cells were then washed with HH buffer and resuspended in HH buffer. The fluorescence intensities of Live cells (healthy, Red) and Dead cells (treated in 55°C water bath for 30 minutes, Green) were measured with NovoCyte 3000 flow cytometer using blue laser APC emission channel. 
Flow Cytometry Analysis of Jurkat cells stained with Calcein Deep Red™ AM ester (Cat#22011). Jurkat cells were washed once with HH buffer and stained with 2 uM Calcein Deep Red™ AM ester (Cat#22011) in HH with 0.02% PF-127(Cat#20053) and 1mM PBC (Cat# 20061) for 30 minutes at 37C incubator. Cells were then washed with HH buffer and resuspended in HH buffer. The fluorescence intensities of Live cells (healthy, Red) and Dead cells (treated in 55°C water bath for 30 minutes, Green) were measured with NovoCyte 3000 flow cytometer using blue laser APC emission channel. 
protocol
Protocol
sds
SDS
CatalogSize
Price
Quantity
220111 mg
Price
 
Physical properties
Molecular weight~1000
SolventDMSO
Spectral properties
Excitation (nm)643
Emission (nm)663
Storage, safety and handling
Intended useResearch Use Only (RUO)
StorageFreeze (< -15 °C); Minimize light exposure
Instrument settings
Flow cytometer
Excitation633/640 nm laser
Emission660/20 nm filter
Instrument specification(s)APC channel
Fluorescence microscope
ExcitationCy5 filter set
EmissionCy5 filter set
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation620
Emission660
Cutoff630
Recommended plateSolid black
Documents
Protocol
Safety Data Sheet
Certificate of Analysis
Flyer: Calcein Dyes
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Page updated on January 23, 2025