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Calcein Violet-AM

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Regulatory Status
RUO
Other Names
Calcein, Calcien Violet-AM
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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
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425203 10 x 25 µg $211
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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
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.
Application

FC - Quality tested
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. 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 Violet-AM dye with 40 µl of anhydrous DMSO.
3. Prepare a 1 µM working solution by diluting 1 µL of 1 mM Calcein Violet-AM stock solution in 1 mL PBS.
4.  Spin down cells and adjust the cell suspension to 1x107 cells/mL in PBS.
5. Add 10 µl of the 1 µM Calcein Violet-AM working solution to each mL of cell suspension for a final concentration of 0.01 µM.
6. Incubate cells for 20 minutes at room temperature or at 37°C, 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 Violet-AM.
9. 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. Fr÷hlich 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)

Antigen Details

Structure
Acetoxymethyl ester.
Distribution

Cytoplasmic.

Function
Cell vitality indicator.
Interaction
Nonspecific esterases.
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Proliferation and Viability
Gene ID
NA

Related FAQs

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Go To Top Version: 3    Revision Date: 02/04/2021

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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