7-AAD | AAT Bioquest

7-AAD

7-Aminoactinomycin D; CAS 7240-37-1

7-AAD (7-aminoactinomycin D) is a cell-impermeant, G-C selective DNA intercalating dye with red fluorescence that specifically stains dead and necrotic cells for viability assessment in flow cytometry and microscopy.

G-C Selective DNA Intercalation: High affinity for GC-rich regions with red fluorescence (Ex/Em = 549/648 nm)
Cell Impermeant Viability Dye: Specifically stains dead and necrotic cells with compromised membranes
488 nm Laser Compatible: Large Stokes shift reduces spectral overlap with FITC and PE in multicolor flow cytometry
Lower Quantum Yield vs PI: QY = 0.02 provides controlled fluorescence intensity for sensitive detection

7-AAD spectrum. 7-AAD (7-Aminoactinomycin D) is a fluorescent compound with an excitation peak at 549 nm and an emission peak at 648 nm. Other spectra of interest include: Propidium iodide, Acridine Orange, and DAPI (4,6-Diamidino-2-phenylindole). 7-AAD (7-Aminoactinomycin D) belongs to the following categories: Cell Cycle Assays, Nucleus, Cell Viability Assays, and Fluorescence Activated Cell Sorting (FACS).

Catalog	Size	Price	Quantity
17501	1 mg	Price

Overview

7-AAD is a cell-impermeant, G-C affinity DNA stain with an excitation peak at 549 nm and an emission peak at 648 nm.

7-aminoactinomycin (7-AAD) is an intercalating dye that binds to double-stranded DNA (dsDNA). It has a strong preference for G-C regions, in contrast to other DNA stains, such as DAPI (A-T affinity) and propidium iodide (no sequence specificity). Because 7-AAD is cell impermeable, it cannot be used for live cell staining, unlike Hoechst 33342 or acridine orange, which are membrane permeable. As such, 7-AAD is most often used to label cells with compromised plasma membranes, such as fixed, necrotic or otherwise nonviable cells, serving as a viability dye for flow cytometry applications.

The most common application of 7-AAD is flow cytometry. In particular, it can be used for cell cycle determination, cell sorting and studying of apoptosis. For apoptosis studies, it is frequently paired with an annexin v conjugate (e.g. Annexin V FITC), wherein annexin v identifies early apoptosis through binding to translocated phosphatidylserine while 7-AAD marks necrotic cells with damaged membranes.

7-AAD vs PI

7-AAD often serves as a replacement for propidium iodide (PI). Both fluorophores are membrane impermeant, staining non-viable or fixed cells. Likewise, both can be excited by the 488 nm argon laser line. One difference is that propidium iodide has a much larger quantum yield (0.2) versus 7-AAD (0.02). This results in a brighter fluorescence signal, on average, for PI. On the other hand, 7-AAD has a significantly large Stokes’ shift. This reduces spectral overlap in multiplex analysis, making 7-AAD more compatible with common fluorophores such as FITC and phycoerythrin (PE).

Excitation and emission

7-AAD has a molecular weight of 1270.43 daltons. It has an excitation wavelength of 549 nm and an emission wavelength of 648 nm. It is well excited by the 488 nm argon laser line. It can be read using a flow cytometer with the PE/Cy5 or FL3 channel.

Preparation of staining solution

A 1 to 10 mM stock solution of 7-AAD can be prepared using DMSO. The stock solution is good for 6 months, if stored in a dark location at < -20 °C.

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.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	78.714 µL	393.568 µL	787.135 µL	3.936 mL	7.871 mL
5 mM	15.743 µL	78.714 µL	157.427 µL	787.135 µL	1.574 mL
10 mM	7.871 µL	39.357 µL	78.714 µL	393.568 µL	787.135 µL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	1270.43 g/mol	=	mL	x	mM	=	mmol

Citations

View all 14 citations: Citation Explorer

Propofol elicits apoptosis and attenuates cell growth in esophageal cancer cell lines

Authors:

Zhou, Rui and Konishi, Yuko and Zhang, Ailing and Nishiwaki, Kimitoshi

Journal:

Nagoya J. Med. Sci (2023): 579--591

Reconstitution and post-thaw storage of cryopreserved human mesenchymal stromal cells: Pitfalls and optimizations for clinically compatible formulants

Authors:

Aabling, Rasmus Roost and Alstrup, Toke and Kj{\ae}r, Emma Mader and Poulsen, Kristine Juul and Pedersen, Jonas Oute and Revenfeld, Anne Louise and M{\o}ller, Bjarne Kuno and Eijken, Marco

Journal:

Regenerative Therapy (2023): 67--75

Rapid fabrication by digital light processing 3D printing of a SlipChip with movable ports for local delivery to ex vivo organ cultures

Authors:

Catterton, Megan and Ball, Alexander and Pompano, Rebecca

Development and improved fabrication of microfluidic devices for local delivery to investigate properties of healthy and inflamed lymph node tissue ex vivo slices.

Authors:

Catterton, Megan Ann

Labelling primary immune cells using bright blue fluorescent nanoparticles

Authors:

Belanger, Maura C and Zhuang, Meng and Ball, Alexander G and Richey, Kristen H and DeRosa, Christopher A and Fraser, Cassandra L and Pompano, Rebecca R

Journal:

Biomaterials science (2020): 1897--1909

References

View all 11 references: Citation Explorer

A study of interaction of 7-aminoactinomycin D with DNA by fluorescence correlation spectroscopy

Authors:

Kovalev AE, Iakovenko AA, Vekshin NL.

Journal:

Biofizika (2004): 1030

Phagocytosis of apoptotic cells assessed by flow cytometry using 7-Aminoactinomycin D

Authors:

Gaforio JJ, Serrano MJ, Algarra I, Ortega E, Alvarez de Cienfuegos G.

Journal:

Cytometry (2002): 8

Multiparametric flow cytometric analysis of biochemical and functional events associated with apoptosis and oncosis using the 7-aminoactinomycin D assay

Authors:

Lecoeur H, de Oliveira-Pinto LM, Gougeon ML.

Journal:

J Immunol Methods (2002): 81

Accessibility of 7-aminoactinomycin D to lymphocyte nuclei after paraformaldehyde fixation

Authors:

Linden E, Skoglund P, Rundquist I.

Journal:

Cytometry (1997): 92

High affinity binding of 7-aminoactinomycin D and 4',6-diamidino-2-phenylindole to human neutrophilic granulocytes and lymphocytes

Authors:

Loborg H, Linden E, Lonn A, Skoglund P, Rundquist I.

Journal:

Cytometry (1995): 296

Physical properties

Molecular weight	1270.43
Solvent	DMSO

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	27500 ¹
Excitation (nm)	549
Emission (nm)	648
Quantum yield	0.02 ¹

Storage, safety and handling

Certificate of Origin	Download PDF
H-phrase	H303, H313, H340
Hazard symbol	T
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R68
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	41116134
CAS	7240-37-1

Documents

Protocol
Safety Data Sheet
Certificate of Origin
Certificate of Analysis

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Page updated on September 24, 2024