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Cat. No.: 7CVAK-200 (for 200T)

Cat. No.: 7CVAK-1k (for 1000T)

Description

7-AAD Cell Viability Assay Kit is a reagent kit developed for the specific staining of necrotic cells based on the characteristics of 7-AAD (7-aminoactinomycin D), a far-red infrared fluorescent probe that stains the cell nucleus of necrotic cells specifically by marking the loss of cell membrane integrity. This kit can be used in conjunction with Annexin V-FITC/EGFP/PE, Calcein AM, and other dyes for the detection of cell apoptosis, necrosis, or cell viability and is suitable for flow cytometry, fluorescence microscopy, or other fluorescence detection devices.

7-AAD, also known as 7-aminoactinomycin D, is a fluorescent indicator that can intercalate with nucleic acids. The DNA complex formed can emit maximum fluorescence at 647nm when excited by 546nm light. This property makes it commonly used in multicolor fluorescence microscopy or flow cytometry experiments.

Similar to propidium iodide (PI), 7-AAD is a non-cell membrane-permeable fluorescent dye that cannot pass through biologically active cytoplasmic membranes. Therefore, it is used to distinguish between normal cells and necrotic cells and can also be used for nuclear staining after cell fixation or permeabilization. Unlike PI, after excitation by a 546nm argon laser, 7-AAD has a narrower emission spectrum and a longer emission wavelength. It has less interference with other detection channels, making it the preferred alternative to PI in multicolor fluorescence analysis. It can be used in combination with various fluorescent dyes excited by 488nm light, such as FITC (fluorescein isothiocyanate), PE (phycoerythrin), APC (allophycocyanin), Calcein AM, etc. It exhibits weak fluorescence around 600nm in the spectrum, which can be detected as red fluorescence under a standard fluorescence microscope, and strong fluorescence at around 650nm in the far-red range, which can be detected using the FL3 channel of a flow cytometer or a fluorescence microscope equipped with a 650nm long-pass filter.

The molecular formula of 7-AAD is C62H87N13O16, with a molecular weight of 1270.45, and a CAS number of 7240-37-1. After intercalation into DNA, its maximum excitation wavelength is 546nm, and the maximum emission wavelength is 647nm.

This assay kit includes a detection buffer that allows for a straightforward and convenient detection of cell necrosis while maintaining the normal cell state.

Components

• 7-AAD (20X)
• Detection Buffer

Storage

Store at -20°C, with a minimum shelf life of one year. 7-AAD (20X) should be stored away from light.

Precaution

• If there is bacterial or fungal contamination, it may seriously impact the effectiveness of the detection.
• It is advisable to perform the test promptly after staining, as an extended period may lead to an increase in the number of necrotic cells.
• After cell fixation, permeabilization, and RNase treatment, 7-AAD can also be used to detect the cell cycle, but propidium iodide (PI) is more commonly employed for this purpose.
• All fluorescent dyes are susceptible to photobleaching, so please make every effort to minimize exposure to light to reduce fluorescence photobleaching.
• This product is intended solely for scientific research by professionals and should not be used for clinical diagnosis or treatment, in food or pharmaceuticals, and should not be stored in residential environments.
• For your safety and health, please wear laboratory attire and disposable gloves while handling.

Related: Annexin V-FITC Apoptosis Detection Kit

Only for research and not intended for treatment of humans or animals

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