Cell apoptosis assay was performed as previously described.26 (link) Briefly, CD19-CAR T cells treated with or without metformin were stained with Annexin V-APC and PI (Biolegend) according to the manufacturer’s instructions. Cells were incubated at room temperature for 20 minutes and then analyzed by BD LSRFortessa™ (BD Biosciences), and data were analyzed by FlowJo software (FlowJo).
Annexin 5 apc and pi
Manufactured by BioLegend
Annexin V-APC and PI is a fluorescent dye combination used to detect apoptosis in cells. Annexin V binds to phosphatidylserine exposed on the cell surface during apoptosis, while propidium iodide (PI) stains the DNA of cells with compromised cell membranes. The combination of these two dyes allows for the identification of viable, early apoptotic, and late apoptotic/necrotic cell populations.
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Lab products found in correlation
2 protocols using annexin 5 apc and pi
1
Apoptosis Assay for CAR T Cells
2
Apoptosis Induction in Cancer Cells
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Cells were seeded in 6-well plates and allowed to adhere for 24 h. Cells were then dosed with doxorubicin (0.5 µM), OKI-005 (0.2 µM), or the combination for 24 and 48 h. Cells were then trypsinized, neutralized with media, and washed with PBS. Cells were transferred to flow cytometry tubes and resuspended with Annexin-V-FITC and PI (Cell Signaling Technology, Danvers, MA, #6592). Due to the GFP-tagged constructs in CAL-51 SCR and CAL-51 p53-10 cell lines, Annexin-V-APC and PI were used (Biolegend, San Diego, CA, #640,919). Samples were run and analyzed by the UCCC Flow Cytometry Shared Resource.
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