Biolegend, 425205, Calcein Red-AM, 10x25μg

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.
10x25μg
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: FC - Quality testedLive cell imaging - Verified
Recommended Usage: 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: 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-AM1 vial of 500 µL anhydrous DMSOCalcein-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.
Structure: Acetoxymethyl ester
Distribution: Cytoplasmic
Function: Cell vitality and health probe. Shows the pathway to death in more detail.
Interaction: Nonspecific esterases
Gene ID: NA
Regulatory Status: RUO
Other Names: Calcein, Calcien Red-AM