7 amino actinomycin d

Manufactured by Southern Biotech

Sourced in United States

7-amino-actinomycin D (7AAD) is a fluorescent dye commonly used in flow cytometry applications. It is a DNA-binding agent that exhibits fluorescence upon intercalation with nucleic acids. 7AAD is used to detect cell viability and identify dead or dying cells in a sample.

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Lab products found in correlation

Q vd oph, by MedChemExpress (1 mentions) Celltiter aqueous one solution cell proliferation assay, by Promega (1 mentions) Facsaria, by BD (1 mentions) Navitoclax, by Active Motif (1 mentions) Venetoclax, by MedChemExpress (1 mentions) Iacs 010759, by Selleck Chemicals (1 mentions)

2 protocols using 7 amino actinomycin d

Annexin V-PE and 7AAD Apoptosis Assay

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To determine the impact of FHC on apoptosis in established cell lines, annexin V-phycoerythrin (PE) and 7-amino-actinomycin D (7AAD) (SouthernBiotech; USA) staining as well as flow cytometry were performed. Briefly, PK-15 cells grown in six-well plates were digested with trypsin and suspended to a concentration of 0.5–1.0 × 10⁶ cells ml⁻¹. Subsequently, 5 μl annexin V-PE were added to 100 μl cells in a new tube, which were incubated on ice for 15 min., followed by sequentially adding 385 μl cold binding buffer and 10 μl 7AAD to each tube, which were incubated on ice for another 15 min. Apoptotic cells were examined using a Coulter Epics XL FACS (Beckman, USA).

Qian G., Lv H., Lin J., Li X., Lv Q., Wang T., Zhang J., Dong W., Guo K, & Zhang Y. (2018). FHC, an NS4B-interacting Protein, Enhances Classical Swine Fever Virus Propagation and Acts Positively in Viral Anti-apoptosis. Scientific Reports, 8, 8318.

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Cell Proliferation and Apoptosis Assay Protocol

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To measure cell proliferation, cells were plated in 96- or 384-well plates (3000 or 1250 cells/well, 3–6 wells/sample) with drugs and incubated for 48 hours at 37°C in 5% CO₂. After incubation, relative numbers of viable cells were measured using a tetrazolium-based colorimetric assay (CellTiter Aqueous One Solution Cell Proliferation Assay, Promega). Inhibitor combination screening was carried out in pre-treated 384 well plates with inhibitors prepared in a 7-point concentration series ranging from 10 μM to 0.014 μM for each drug (23 (link)).
To measure cell apoptosis, cells (in duplicates) were resuspended in 100 μL of Annexin V binding buffer containing 1 μL of Annexin V-PE, 1 μL of 7-aminoactinomycin D (7-AAD) (Southern Biotech) and 1 μL of CD19-mAbs (BD Bioscience) followed by flow cytometry on FACSAria (BD Biosciences). Data analysis was performed using FlowJo software (Tree Star).
AZD5991 and navitoclax were purchased from Active Biochem; venetoclax and Q-VD-OPh were from MedChemExpress; AZD4320, IACS-010759 were from Selleck Chemicals. CCCP (carbonyl cyanide m-chlorophenyl hydrazine), NAC (N-Acetyl-L-cysteine) were from Sigma.

Liu T., Lam V., Thieme E., Sun D., Wang X., Xu F., Wang L., Danilova O.V., Xia Z., Tyner J.W., Kurtz S.E, & Danilov A.V. (2021). Pharmacologic targeting of Mcl-1 induces mitochondrial dysfunction and apoptosis in B-cell lymphoma cells in a TP53- and BAX-dependent manner. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(17), 4910-4922.

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