Calcein-AM

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Other Names
Calcein, Calcein AM
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Calcein-AM_FC_1_080216
Fresh (top) or day-old C57BL/6 splenocytes (bottom) were stained with Calcein-AM and a cell-impermeant nucleic acid dye, SYTOX™ Red (colored). Black figure represents unstained splenocytes.
  • Calcein-AM_FC_1_080216
    Fresh (top) or day-old C57BL/6 splenocytes (bottom) were stained with Calcein-AM and a cell-impermeant nucleic acid dye, SYTOX™ Red (colored). Black figure represents unstained splenocytes.
  • Calcein-AM_FC_2_080216
Cat # Size Price Quantity Avail. Save
425201 10 x 50 µg $140
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Description

Calcein-AM is a fluorogenic, cell-permeant fluorescent probe that indicates cellular health. When the acetoxymethyl ester is intact, this probe is nonfluorescent until acted upon by nonspecific esterases present within the healthy, live cells. Cleaving the AM ester allows the probe to excite and emit at 488nm/ 520nm respectively. The signal of Calcein-AM is proportional to cell vitality, as esterase activity decreases in cells with poor vitality. This cell-permeant probe is not retained with fixation and peameabilization.

Product Details
Technical data sheet

Product Details

Preparation
Calcein-AM labeling kit consists of lyophilized Calcein-AM and anhydrous DMSO. For reconstitution, bring the kit to room temperature and add 50 µL of DMSO to one vial of Calcein-AM dye until fully dissolved.
Storage & Handling
Upon receipt, Calcein-AM should be stored at -20°C. Do not open vials until needed. Once the DMSO is added to the Calcein-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-AM is 994 Da. The maximum excitation and emission wavelengths of Calcein-AM are 488 nm/520 nm, respectively. Each 50 µg of Calcein-AM may be reconstituted with 50 µL of anhydrous DMSO to yield a stock concentration of 1 mM.

Materials Provided:
10 vials x 50 µg Calcein-AM
2 vials 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-AM dye with 50 µl of anhydrous DMSO.
3. Prepare a 1 µM working solution by diluting 1 µL of 1 mM Calcein-AM stock solution in 1 mL PBS.
4. Spin down cells and adjust the cell suspension to 1x107 cells/mL in PBS.
5. Add 10µl of the 1 µM Calcein-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 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-AM.
9. After incubation, Calcein-AM labeled cells are ready for downstream applications or analysis.

Application References

(PubMed link indicates BioLegend citation)
  1. Mariappan MR, et al. 1999. IEEE Eng. Med. Biol. Mag. 18:22.
  2. Mironov SL, et al. 2005. J. Biol. Chem. 280:715.
  3. Tian F, et al. 2009. Blood. 113:5352.
  4. Ansbro MR, et al. 2013. PLoS One 8(4):e60334. (IF)
  5. Gonzalez JM, et al. 2013. Invest. Ophthalmol. Vis. Sci. 54:1039. (IF)
  6. Godin LM, et al. 2011. Am. J. Physiol. Lung Cell Mol. Physiol. 300:L615. (IF)

Antigen Details

Structure
Acetoxymethyl ester.
Distribution

Cytoplasmic.

Function
Cell vitality indicator.
Interaction
Nonspecific esterases.
Biology Area
Cell Proliferation and Viability
Gene ID
NA

Related FAQs

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Go To Top Version: 2    Revision Date: 08/17/2016

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