BD Via-ProbeTM Red Nucleic Acid Stain

Description

Panel 1 - Flow cytometric analysis of HeLa cell viability. Cells from the human HeLa Cervical adenocarcinoma, ATCC CCL 2 cell line were heat-killed by incubation at 60C 45 min , cooled to room temperature, and mixed with an equal number of untreated viable HeLa cells. Cells were then stained with BD Via-Probe Red Nucleic Acid Stain Cat. No. 565803/565804; 10 nM and then acquired by flow cytometry. Morphologically live red solid line histogram and dead blue dashed line histogram HeLa cells were gated based on their distinct forward and side light-scattering characteristics and can be clearly distinguished based on staining intensity of BD Via-Probe Red. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 2 - Two-color flow cytometric analysis of mouse thymocyte viability. BALB/c thymocytes were left untreated Left Plot or stimulated with Purified NA/LE Hamster Anti-Mouse CD95 antibody Cat. No. 554254; Right Plot for 4 hours. Cells were harvested, stained with BD Horizon BV421 Annexin V Cat. No. 563973 and BD Via-Probe Red Nucleic Acid Stain 5 nM , and analyzed by flow cytometry. Anti-CD95-stimulated cells show an increase in Annexin V-positive and BD Via-Probe Red-positive cells, indicating an increase in apoptosis and cell death. Flow cytometric analysis was performed using a BD LSRFortessa X-20 Cell Analyzer System. Panel 1 - Flow cytometric analysis of HeLa cell DNA content. HeLa cells in log phase growth were harvested, fixed, and permeabilized using 70 % ice-cold ethanol while vortexing. Cells were resuspended in DPBS with 0.1 muM BD Via-Probe Red Nucleic Acid Stain and 0.25 mug/mL RNAse A Sigma, Cat. No. R6513 and acquired by flow cytometry at a low flow rate using a BD LSRFortessa Cell Analyzer System. DNA histograms were deconvoluted by FlowJo software into G0/G1, S, and G2/M populations. Please note that for DNA content analysis, this dye is also compatible with BD Pharmingen Transcription Factor Buffer Set Cat. No. 562574/562725 . Panel 2 - Immunofluorescent staining of HeLa cells. HeLa cells were seeded into a 96-well imaging plate at 10,000 cells/well. After overnight incubation, the cells were fixed in BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 , and then stained with Alexa Fluor 488 Mouse Anti-Actin antibody Cat. No. 558623; pseudocolored green . Cell nuclei were counterstained with BD Via-Probe Red Nucleic Acid Stain pseudocolored red with 0.25 mg/mL RNAse A Sigma Aldrich, Cat. No. R6513 . The images were captured on a Molecular Devices ImageXpress Micro XLS using a 20 objective and merged using Molecular Devices MetaXpress software. Red Nucleic Acid Stain is also compatible with Saponin and Triton X-100 fix/perm protocols. Panel 1 - Flow cytometric analysis of HeLa cell viability. Cells from the human HeLa Cervical adenocarcinoma, ATCC CCL 2 cell line were heat-killed by incubation at 60C 45 min , cooled to room temperature, and mixed with an equal number of untreated viable HeLa cells. Cells were then stained with BD Via-Probe Red Nucleic Acid Stain Cat. No. 565803/565804; 10 nM and then acquired by flow cytometry. Morphologically live red solid line histogram and dead blue dashed line histogram HeLa cells were gated based on their distinct forward and side light-scattering characteristics and can be clearly distinguished based on staining intensity of BD Via-Probe Red. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 2 - Two-color flow cytometric analysis of mouse thymocyte viability. BALB/c thymocytes were left untreated Left Plot or stimulated with Purified NA/LE Hamster Anti-Mouse CD95 antibody Cat. No. 554254; Right Plot for 4 hours. Cells were harvested, stained with BD Horizon BV421 Annexin V Cat. No. 563973 and BD Via-Probe Red Nucleic Acid Stain 5 nM , and analyzed by flow cytometry. Anti-CD95-stimulated cells show an increase in Annexin V-positive and BD Via-Probe Red-positive cells, indicating an increase in apoptosis and cell death. Flow cytometric analysis was performed using a BD LSRFortessa X-20 Cell Analyzer System. Show More Panel 1 - Flow cytometric analysis of HeLa cell DNA content. HeLa cells in log phase growth were harvested, fixed, and permeabilized using 70 % ice-cold ethanol while vortexing. Cells were resuspended in DPBS with 0.1 muM BD Via-Probe Red Nucleic Acid Stain and 0.25 mug/mL RNAse A Sigma, Cat. No. R6513 and acquired by flow cytometry at a low flow rate using a BD LSRFortessa Cell Analyzer System. DNA histograms were deconvoluted by FlowJo software into G0/G1, S, and G2/M populations. Please note that for DNA content analysis, this dye is also compatible with BD Pharmingen Transcription Factor Buffer Set Cat. No. 562574/562725 . Panel 2 - Immunofluorescent staining of HeLa cells. HeLa cells were seeded into a 96-well imaging plate at 10,000 cells/well. After overnight incubation, the cells were fixed in BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 , and then stained with Alexa Fluor 488 Mouse Anti-Actin antibody Cat. No. 558623; pseudocolored green . Cell nuclei were counterstained with BD Via-Probe Red Nucleic Acid Stain pseudocolored red with 0.25 mg/mL RNAse A Sigma Aldrich, Cat. No. R6513 . The images were captured on a Molecular Devices ImageXpress Micro XLS using a 20 objective and merged using Molecular Devices MetaXpress software. Red Nucleic Acid Stain is also compatible with Saponin and Triton X-100 fix/perm protocols. Show More Panel 1 - Flow cytometric analysis of HeLa cell viability. Cells from the human HeLa Cervical adenocarcinoma, ATCC CCL 2 cell line were heat-killed by incubation at 60C 45 min , cooled to room temperature, and mixed with an equal number of untreated viable HeLa cells. Cells were then stained with BD Via-Probe Red Nucleic Acid Stain Cat. No. 565803/565804; 10 nM and then acquired by flow cytometry. Morphologically live red solid line histogram and dead blue dashed line histogram HeLa cells were gated based on their distinct forward and side light-scattering characteristics and can be clearly distinguished based on staining intensity of BD Via-Probe Red. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 2 - Two-color flow cytometric analysis of mouse thymocyte viability. BALB/c thymocytes were left untreated Left Plot or stimulated with Purified NA/LE Hamster Anti-Mouse CD95 antibody Cat. No. 554254; Right Plot for 4 hours. Cells were harvested, stained with BD Horizon BV421 Annexin V Cat. No. 563973 and BD Via-Probe Red Nucleic Acid Stain 5 nM , and analyzed by flow cytometry. Anti-CD95-stimulated cells show an increase in Annexin V-positive and BD Via-Probe Red-positive cells, indicating an increase in apoptosis and cell death. Flow cytometric analysis was performed using a BD LSRFortessa X-20 Cell Analyzer System. Panel 1 - Flow cytometric analysis of HeLa cell DNA content. HeLa cells in log phase growth were harvested, fixed, and permeabilized using 70 % ice-cold ethanol while vortexing. Cells were resuspended in DPBS with 0.1 muM BD Via-Probe Red Nucleic Acid Stain and 0.25 mug/mL RNAse A Sigma, Cat. No. R6513 and acquired by flow cytometry at a low flow rate using a BD LSRFortessa Cell Analyzer System. DNA histograms were deconvoluted by FlowJo software into G0/G1, S, and G2/M populations. Please note that for DNA content analysis, this dye is also compatible with BD Pharmingen Transcription Factor Buffer Set Cat. No. 562574/562725 . Panel 2 - Immunofluorescent staining of HeLa cells. HeLa cells were seeded into a 96-well imaging plate at 10,000 cells/well. After overnight incubation, the cells were fixed in BD Cytofix Fixation Buffer Cat. No. 554655 , permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 , and then stained with Alexa Fluor 488 Mouse Anti-Actin antibody Cat. No. 558623; pseudocolored green . Cell nuclei were counterstained with BD Via-Probe Red Nucleic Acid Stain pseudocolored red with 0.25 mg/mL RNAse A Sigma Aldrich, Cat. No. R6513 . The images were captured on a Molecular Devices ImageXpress Micro XLS using a 20 objective and merged using Molecular Devices MetaXpress software. Red Nucleic Acid Stain is also compatible with Saponin and Triton X-100 fix/perm protocols.