Calcein Violet-AM

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Other Names
CFDA-SE, CFSE, cell tracking
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Calcein_Violet_AM_1_093016
Fresh (top) or day-old C57BL/6 splenocytes (bottom) were stained with 0.01 µM Calcein Violet-AM and a cell-impermeant nucleic acid dye, SYTOX™ Red (colored). Black figure represents unstained splenocytes.
  • Calcein_Violet_AM_1_093016
    Fresh (top) or day-old C57BL/6 splenocytes (bottom) were stained with 0.01 µM Calcein Violet-AM and a cell-impermeant nucleic acid dye, SYTOX™ Red (colored). Black figure represents unstained splenocytes.
  • Calcein_Violet_AM_2_093016
Description

Calcein Violet-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 400nm/452nm respectively. The signal of Calcein Violet-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
Cat # Size Price Quantity Avail. Save
425203 10 x 25 µg $180.00
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Technical data sheet

Product Details

Preparation
Calcein Violet-AM labeling kit consists of lyophilized Calcein Violet-AM and anhydrous DMSO. For reconstitution, bring the kit to room temperature and add 40 µL of DMSO to one vial of Calcein Violet-AM dye until fully dissolved.
Storage & Handling
Calcein Violet-AM should be stored at -20°C upon receipt. Do not open vials until needed. Once the DMSO is added to the Calcein Violet-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 one month. The unopened kit stored at -20°C is guaranteed for one year after purchase.
Application

FC - Quality tested
IF - Reported in the literature

Recommended Usage

This lot has been tested by flow cytometric analysis of cell vitality. It can be used at concentrations ranging from 0.1 – 0.001 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 Violet-AM is 616.48 Da. The maximum excitation and emission wavelengths of Calcein Violet-AM are 400 nm/452 nm, respectively. Each 25 µg of Calcein Violet-AM may be reconstituted with 40 µL of anhydrous DMSO to yield a stock concentration of 1 mM.

Materials Provided:
10 vials x 25 µg Calcein Violet-AM
2 vials of 500 ul anhydrous DMSO

Calcein Violet-AM Labeling Procedure:
1. Prepare a 1 mM stock solution by reconstituting 1 vial of lyophilized Calcein Violet-AM dye with 40 µl of anhydrous DMSO.
2. Prepare a 1 µM working solution by diluting 1 µL of 1 mM Calcein Violet-AM stock solution in 1 mL PBS.
3.  Spin down cells and adjust the cell suspension to 1x107 cells/mL in PBS.
4. Add 10ml of the 1mM Calcein Violet-AM working solution to each mL of cell suspension for a final concentration of 0.01mM.
5. Incubate cells for 20 minutes at room temperature or at 37°C, protected from light.
6. Pellet cells and resuspend in pre-warmed cell culture medium.
7. Incubate cells for 10 minutes to ensure optimal retention of the Calcein Violet-AM.
8. After incubation, Calcein Violet-AM labeled cells are ready for downstream applications or analysis.

Application References

(PubMed link indicates BioLegend citation)

1. Eastburn DJ, et al. 2014. Nucleic Acids Res. 42:e128. (FC)
2. Zhou Q, et al. 2012. Blood. 120:4334. (FC)
3. Fröhlich JD, et al. 2012. Am. J. Pathol. 180:153.
4. Wang T, et al. 2012. ACS Nano. 6:1251.
5. Lis R, et al. 2011. Int. J. Cancer 128:715. (FC)
6. Cole LE, et al. 2007. Infect. Immun. 75:4127. (IF)

Publication Library

Antigen Details

Structure
Acetoxymethyl ester.
Distribution
Cytoplasmic.
Function
Cell vitality indicator.
Interaction
Nonspecific esterases.
Gene ID
NA

Related FAQs

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Go To Top Version: 1    Revision Date: 09/30/2016

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