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Calcein Red-AM

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Regulatory Status
RUO
Other Names
Calcein, Calcien Red-AM
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Calcein-Red_AM_032720.png
Non-treated Jurkats (left) or treated Jurkats (right) that have been stimulated with BioLegend’s anti-human CD95 (EOS9.1) for four hours were then stained with 0.006 µM Calcein Red-AM and 5µL of Annexin Brilliant Violet 421™ (targets and identifies apoptotic cells).
  • Calcein-Red_AM_032720.png
    Non-treated Jurkats (left) or treated Jurkats (right) that have been stimulated with BioLegend’s anti-human CD95 (EOS9.1) for four hours were then stained with 0.006 µM Calcein Red-AM and 5µL of Annexin Brilliant Violet 421™ (targets and identifies apoptotic cells).
See Calcein Red-AM spectral data
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425205 10 x 25 µg 211 CHF
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Description

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.

Product Details
Technical Data Sheet (pdf)

Product Details

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 tested
Live 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-AM
1 vial 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 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.

Antigen Details

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
Go To Top Version: 1    Revision Date: 03.27.2020

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.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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