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Calcein-AM

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Regulatory Status
RUO
Other Names
Calcein, Calcein AM
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Product Citations
publications
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
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425201 10 x 50 µg $165
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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.
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-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)
Product Citations
  1. Bhatt RS, et al. 2021. Cancer Immunol Res. 9:156. PubMed
  2. Yongabi D, et al. 2022. Adv Sci (Weinh). 9:e2200459. PubMed
  3. Oude Egberink R, et al. 2022. Pharmaceutics. 14: . PubMed
  4. McNulty J, et al. 2023. Sci Rep. 13:1639. PubMed
  5. Li S, et al. 2022. Elife. 11:. PubMed
  6. Abeynaike S, et al. 2023. Viruses. 15:. PubMed
  7. Yang F, et al. 2022. J Cell Mol Med. 26:2363. PubMed
  8. Chan JM, et al. 2021. Cancer Cell. 39:1479. PubMed
  9. Deng J, et al. 2021. Breast Cancer Res Treat. 185:85. PubMed
  10. Wang K, et al. 2020. Sci Adv. 6:eaba7606. PubMed
  11. Quintanal-Villalonga A, et al. 2022. STAR Protoc. 3:101776. PubMed
  12. Riedelberger M, et al. 2020. Bio Protoc. e3552:10. PubMed
  13. Huang RS, et al. 2021. Curr Protoc. 1:e246. PubMed
  14. Huang N, et al. 2020. Genome Biol. 1.03125. PubMed
  15. Lavoie RR, et al. 2021. Cancers (Basel). 13:. PubMed
  16. Wachendörfer M, et al. 2022. Biomater Sci. :. PubMed
  17. Hedemann N, et al. 2021. Cancers (Basel). 13:. PubMed
  18. Feng Y, et al. 2022. Life (Basel). 12:. PubMed
  19. Rossignol A, et al. 2017. MAbs. 9:521. PubMed

Antigen Details

Structure
Acetoxymethyl ester.
Distribution

Cytoplasmic.

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