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7aad via probe

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The 7AAD Via Probe is a fluorescent dye used for the detection of dead cells in flow cytometry applications. It binds to DNA, allowing the identification of non-viable cells.

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Lab products found in correlation

Lsrii flow cytometer, by BD (1 mentions) Anti cd14 percp, by BD (1 mentions) Anti cd3 pe, by BD (1 mentions) Anti cd11c apc, by BD (1 mentions) Anti cd80 pe, by BD (1 mentions) Anti cd66b pe, by BD (1 mentions) Cd34 apc, by BD (1 mentions) Draq5, by Cell Signaling Technology (1 mentions) Cd49d fitc, by Miltenyi Biotec (1 mentions)

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Via-Probe (47 citations)

4 protocols using 7aad via probe

1

Quantifying Cell Death in HCT116 Cells

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Following BK and NT analogue treatment of HCT116 cells, we evaluated the proportion of dead cells in our samples. For this purpose, we used a 7AAD Via Probe (BD Biosciences, San Jose, CA, USA). The cells were incubated for 30 min with 10 µm of a dye, washed with PBS, and prepared for FACS analysis. We used unstained cells to set a threshold for positive signal. The frequency of 7AAD-positive cells was compared to untreated control cells.
Szaryńska M., Olejniczak-Kęder A., Podpłońska K., Prahl A, & Iłowska E. (2023). Bradykinin and Neurotensin Analogues as Potential Compounds in Colon Cancer Therapy. International Journal of Molecular Sciences, 24(11), 9644.
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2

Evaluating Cell Death in CRC Cells

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Following the T3 treatment of HCT116 and HT29 cells and cancer cells isolated from CRC patients the proportion of dead cells in our samples was evaluated. For this purpose, 7AAD Via Probe (BD Biosciences) was used. The cells were incubated for 30 min at RT with 10 µm of a dye, washed with PBS and prepared for FACS analysis. To set a threshold of positive signal, unstained cells were used. The frequency of 7AAD− positive cells was compared with untreated control cells.
Rostkowska O., Olejniczak-Kęder A., Spychalski P., Szaryńska M, & Kobiela J. (2022). Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro. Oncology Reports, 49(1), 21.
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3

Multiparametric Flow Cytometry Analysis

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Differentiating cells during each passage and after stimulation were stained with the following cocktail of monoclonal antibodies purchased from BD Biosciences: anti-CD3-PE, anti-CD14-PerCP, anti-CD66b-PE, anti-CD16-PerCP-Cy5.5, anti-CD11c-APC, anti-CD80-PE, anti-HLA-DR-PerCP, and matched isotype controls. After 30 min of incubation in the dark, on-ice samples were fixed and prepared for further analysis. Flow cytometric analysis was performed using an LSRII flow cytometer (BD). For death cell evaluation a 7AAD Via Probe (BD) was used. After adding 10 μm of Via Probe samples were incubated for 30 minutes, washed, and resuspended in PBS prior to cytometric analysis.
Szaryńska M., Preis K., Zabul P, & Kmieć Z. (2018). Diversity of dendritic cells generated from umbilical cord or adult peripheral blood precursors. Central-European Journal of Immunology, 43(3), 306-313.
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4

Flow Cytometric Analysis of Hematopoietic and Erythroid Cell Differentiation

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For flow cytometry analysis of HSPCs and erythroblasts differentiated from human iPSCs, we harvested ~200,000 cells, washed with PBS, and re-suspended the cell pellet in PBS+1%FBS for antibody staining. We used CD34-APC (BD Biosciences), CD45-AF488 or CD45-APC (Invitrogen), CD235a-FITC (Invitrogen), Band3-FITC or Band3-APC (kindly provided by Dr. Xiuli An at NYBC), CD49d-FITC or CD49d-PE (Miltenyi Biotec, San Diego, CA) to characterize expanded and differentiated cells. Dead cells were excluded from analysis by positive staining 7-AAD (Viaprobe, BD Biosciences).
For flow cytometric analysis of erythroid terminal maturation of erythroblasts, we harvested suspension cells from terminal maturation culture at different time points of maturation, washed with PBS, and re-suspended in PBS+1%FBS and stained with appropriate combinations of CD235a-FITC, Band3-FITC or APC, CD45-APC and CD49d-FITC (BD BioSciences and Life Technologies), and DRAQ5, a cell permeable dye for DNA staining (Cell Signaling Technology, Danvers, MA). Intracellular hemoglobin staining was done as described before [21 (link)].
Huang X., Wang Y., Yan W., Smith C., Ye Z., Wang J., Gao Y., Mendelsohn L, & Cheng L. (2015). Production of gene-corrected adult beta globin protein in human erythrocytes differentiated from patient iPSCs after genome editing of the sickle point mutation. Stem cells (Dayton, Ohio), 33(5), 1470-1479.
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