PE anti-human CD96 (TACTILE) Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Cells were stained with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) (BD Pharmingen, San Diego, CA, USA) to detect the apoptosis state in accordance with the manufacturer specifications.	The cells (1 × 106) were collected and resuspended in 100 μl Cell Staining Buffer, the antibody was added and incubated at 4 °C for 15 min, washed again with cell staining buffer one time, and then subject to the apoptosis assay. To exclude non-viable cells, cells were stained with 7AAD before flow cytometry (BD Versa, San Diego, CA, USA) analysis for antigen detecting.

Protocol tips
Cells were stained with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) (BD Pharmingen, San Diego, CA, USA) to detect the apoptosis state in accordance with the manufacturer specifications.
Downstream tips
The cells (1 × 106) were collected and resuspended in 100 μl Cell Staining Buffer, the antibody was added and incubated at 4 °C for 15 min, washed again with cell staining buffer one time, and then subject to the apoptosis assay. To exclude non-viable cells, cells were stained with 7AAD before flow cytometry (BD Versa, San Diego, CA, USA) analysis for antigen detecting.

Publication protocol

Cell cycle, cell apoptosis, and cell surface antigens were detected using flow cytometry. In brief, 48 h after treatment of KG-1, Kasumi-1, KG-1a, TF-1, or primary cells, the cell density was adjusted according to the cell viability assay and prepared for cell cycle analysis as described in our previous report [17]. Cells were stained with Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) (BD Pharmingen, San Diego, CA, USA) to detect the apoptosis state in accordance with the manufacturer specifications. The CD34+/CD38− population (Kasumi-1 cells) or CD34+/CD38−/CD96+ population (KG-1 cells) was obtained by incubating with CD34–FITC (Biolegend, San Diego, CA USA), CD38–R-phycoerythrin cyanine7 (CD38–PE-Cy™7; Biolegend), and CD96–R-phycoerythrin (CD96-PE; Biolegend) in Cell Staining Buffer (Biolegend); Annexin V–FITC and CD38–PE-Cy™7 were used for KG-1, KG-a, and TF-1 cells, and CD34–APC was also added for primary cells. The procedure was as follows: The cells (1 × 106) were collected and resuspended in 100 μl Cell Staining Buffer, the antibody was added and incubated at 4 °C for 15 min, washed again with cell staining buffer one time, and then subject to the apoptosis assay. To exclude non-viable cells, cells were stained with 7AAD before flow cytometry (BD Versa, San Diego, CA, USA) analysis for antigen detecting.

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