- Regulatory Status
- RUO
- Other Names
- Calcein, Calcien Red-AM
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Non-treated Jurkats (left) or treated Jurkats (right) that have been stimulated with BioLegend’s anti-human CD95 (EOS9.1) for four hours were then stained with 0.006 µM Calcein Red-AM and 5µL of Annexin Brilliant Violet 421™ (targets and identifies apoptotic cells).
| Cat # | Size | Price | Quantity Check Availability | ||
|---|---|---|---|---|---|
| 425205 | 10 x 25 µg | $211.00 | |||
Calcein Red-AM is a fluorogenic, cell-permeant fluorescent probe that indicates cellular health by detecting the activity of nonspecific esterases. When the acetoxymethyl ester is intact the probe is non-fluorescent, but when it is cleaved by nonspecific esterases present in healthy cells, the probe can be excited and emit at 560 nm/574 nm respectively. The signal of Calcein Red-AM is proportional to the cell vitality, as esterase activity decreases in cells with poor vitality. This cell-permeant probe is not retained with fixation and permeabilization.
Product Details
- Formulation
- Lyophilized
- Preparation
- Calcein Red-AM labeling kit consists of lyophilized Calcein Red-AM and anhydrous DMSO. For reconstitution, bring the kit to room temperature and add 25 µL of DMSO to one vial of Calcein Red-AM dye until fully dissolved.
- Storage & Handling
- Calcein Red-AM should be stored at -20°C upon receipt. Do not open vials until needed. Once the DMSO is added to the Calcein Red-AM, use immediately or store at -20°C in a dry place and protected from light, preferably in a desiccator or in a container with desiccant for no more than a month.
- Application
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FC - Quality tested
Live cell imaging - Verified - Recommended Usage
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This lot has been tested by flow cytometric analysis of cell vitality. It can be used at concentrations ranging from 0.025 – 0.006 µM for cell labeling. It is recommended that the reagent be titrated for optimal performance for each cell type, culturing condition, or application.
- Application Notes
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The molecular weight of Calcein Red-AM is 1015.79 Da. The maximum excitation and emission wavelengths of Calcein Red-AM are 560 nm/574 nm, respectively. Each 25 µg of Calcein-AM may be reconstituted with 25 µL of anhydrous DMSO to yield a stock concentration of 1 mM.
Materials Provided:
10 vials x 25 µg Calcein Red-AM
1 vial of 500 µL anhydrous DMSO
Calcein-AM Labeling Procedure:
1. Prior to reconstitution, spin down the vial of lyophilized reagent in a microcentrofuge to ensure the reagent is at the bottom of the vial.
2. Prepare a 1 mM stock solution by reconstituting 1 vial of lyophilized Calcein Red-AM dye with 25 µL of anhydrous DMSO.
3. Prepare a 1 µM working solution by diluting 1 µL of 1 mM Calcein Red-AM stock solution in 1 mL PBS.
4. Spin down cells and adjust the cell suspension to 1x107 cells/mL in PBS.
5. Add 6 µL of the 1 µM Calcein Red-AM working solution to each mL of cell suspension for a final concentration of 0.006 µM.
6. Incubate cells for 30 minutes at room temperature or at 37°C and keep protected from light.
7. Pellet cells and resuspend in pre-warmed cell culture medium.
8. Incubate cells for 10 minutes to ensure optimal retention of the Calcein Red-AM.
9. After incubation, Calcein Red-AM labeled cells are ready for downstream applications or analysis.
Antigen Details
- Structure
- Acetoxymethyl ester
- Distribution
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Cytoplasmic
- Function
- Cell vitality and health probe. Shows the pathway to death in more detail.
- Interaction
- Nonspecific esterases
- Gene ID
- NA