7-AAD [7-Aminoactinomycin D] *CAS 7240-37-1*

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Chemical structure for 7-AAD [7-Aminoactinomycin D] *CAS 7240-37-1*
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1 mg 17501 $75


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Overview

Ex/Em (nm)547/648
MW1270.43
CAS #7240-37-1
SolventDMSO
Storage Freeze (<-15 °C)
Minimize light exposure
Category Cell Viability
7-Amino actinomycin D (7-AAD) is a non-permeant dye that can be used to identify non-viable cells. Cells with damaged plasma membranes or with impaired/no cell metabolism are unable to prevent the dye from entering the cell. Once inside the cell, the dyes bind to intracellular DNA producing highly fluorescent adducts which identify the cells as "non-viable". It is widely used in flow cytometry. 7-AAD is excited by the 488 nm laser line of an argon laser with fluorescence detected above 650 nm. Although the emission intensity of 7-AAD is lower than that of PI, the longer wavelength emission may make it more useful for multiplexing assays in combination with other 488 nm-excited fluorochromes such as FITC and PE.




Calculators
Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of 7-AAD [7-Aminoactinomycin D] *CAS 7240-37-1* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.



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Table 2. Enter any two values (mass, volume, concentration) to calculate the third.

Mass Molecular weight Volume Concentration Moles
/ = x =
 






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Protocol


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This protocol only provides a guideline, and should be modified according to your specific needs.

7-AAD staining is normally performed after all other stainings. It stains dead cells only. The following procedure can be adapted for most cell types. Growth medium, cell density, the presence of other cell types and other factors may influence staining. Residual detergent on glassware may also affect real or apparent staining of many organisms, causing brightly stained material to appear in solutions with or without cells present.

1). Make 1-10 mM DMSO stock solution. The DMSO stock solution is good for 6 months if stored at -20°C.

2). Use the fixation protocol appropriate for your sample.

3). Pellet cells by centrifugation and resuspend the cells in buffered salt solutions or media, with optimal dye staining at pH 7.4. Adherent cells in culture may be stained in situ on cover slips or in the cell culture wells.

4). Add 7-AAD stain using the concentrations from 0.5 to 5 ┬ÁM and incubate it for 15 to 60 minutes as a guide.

Note: In initial experiments, it may be best to try several dye concentrations over the entire suggested range to determine the concentration that yields optimal staining.

5). Measure the fluorescence intensity at Ex/Em = 545/650nm (use FL3 for flow cytometric analysis)

CAUTION: 7-AAD is a potential carcinogen. It is recommended that the user wear gloves, protective clothing, and eye/face protection in order to avoid contact with skin and eyes.






References

PD-1 silencing impairs the anti-tumor function of chimeric antigen receptor modified T cells by inhibiting proliferation activity
Authors: Jianshu Wei, Can Luo, Yao Wang, Yelei Guo, Hanren Dai, Chuan Tong, Dongdong Ti, Zhiqiang Wu, Weidong Han
Journal: Journal for immunotherapy of cancer (2019): 1--15

Resources-Application Notes Phenotypic and Epigenetic Mechanism of Action Determinations of Histone Methylase and Demethylase Inhibitors using Digital Widefield Microscopy
Authors: Brad Larson, Peter Banks
Journal: Experimental Biology (2017)