BD PharmingenTM DAPI Solution

Description

Panel 1. Two-color flow cytometric analysis of Jurkat cell viability. Jurkat cells were treated with DMSO vehicle Left Plot or 5 muM Camptothecin Right Plot overnight. Cells were resuspended in Annexin V Binding Buffer Cat. No. 556454 and stained with FITC Annexin V Cat. No. 556419 and 0.06 mug/mL DAPI. Camptothecin-treated cells show an increased frequency of apoptotic Annexin V+DAPI- and dead Annexin V+DAPI+ cells. Analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 2. Flow cytometric analysis of HeLa cell DNA content. Cultured HeLa cells in log phase growth were harvested using Gibco Cell Dissociation Buffer Life Technologies , fixed, and permeabilized using BD Pharmingen Transcription Factor Buffer Set Cat. No. 562574/562725 . Cells were resuspended in DPBS with 1 mug/mL DAPI and analyzed using a low BD LSRFortessa cytometer flow rate. A 405 nm laser with a 450/50 nm bandpass filter was used to collect data; comparable results were obtained using a 355 nm laser with a 450/50 nm bandpass filter not shown . Histograms were deconvoluted by FlowJo software into G0/G1, S, and G2/M populations. Panel 3. Multicolor immunofluorescence analysis of NKX2.2 and PTF1A expression in mouse pancreas. Following antigen retrieval with BD Retrievagen A Buffer Cat. No. 550524 , a formalin-fixed paraffin-embedded pancreas tissue section was stained with Purified Mouse Anti-Nkx2.2 antibody Cat. No. 564731, pseudo-colored red , washed, and stained with Alexa Fluor 488 Goat Anti-Mouse Ig Life Technologies . After washing, the section was stained with Alexa Fluor 647 Mouse Anti-PTF1A pseudo-colored green and counterstained with DAPI pseudo-colored blue . The nuclei of islet cells DAPI+ stain positively for both NKX2.2 and PTF1A. The nuclei of acinar cells and some islet precursor cells stain positive for PTF1A. All images were analyzed using a BD Pathway 435 High-Content Bioimager System and merged with BD Attovision Software. Panel 1. Two-color flow cytometric analysis of Jurkat cell viability. Jurkat cells were treated with DMSO vehicle Left Plot or 5 muM Camptothecin Right Plot overnight. Cells were resuspended in Annexin V Binding Buffer Cat. No. 556454 and stained with FITC Annexin V Cat. No. 556419 and 0.06 mug/mL DAPI. Camptothecin-treated cells show an increased frequency of apoptotic Annexin V+DAPI- and dead Annexin V+DAPI+ cells. Analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 2. Flow cytometric analysis of HeLa cell DNA content. Cultured HeLa cells in log phase growth were harvested using Gibco Cell Dissociation Buffer Life Technologies , fixed, and permeabilized using BD Pharmingen Transcription Factor Buffer Set Cat. No. 562574/562725 . Cells were resuspended in DPBS with 1 mug/mL DAPI and analyzed using a low BD LSRFortessa cytometer flow rate. A 405 nm laser with a 450/50 nm bandpass filter was used to collect data; comparable results were obtained using a 355 nm laser with a 450/50 nm bandpass filter not shown . Histograms were deconvoluted by FlowJo software into G0/G1, S, and G2/M populations. Panel 3. Multicolor immunofluorescence analysis of NKX2.2 and PTF1A expression in mouse pancreas. Following antigen retrieval with BD Retrievagen A Buffer Cat. No. 550524 , a formalin-fixed paraffin-embedded pancreas tissue section was stained with Purified Mouse Anti-Nkx2.2 antibody Cat. No. 564731, pseudo-colored red , washed, and stained with Alexa Fluor 488 Goat Anti-Mouse Ig Life Technologies . After washing, the section was stained with Alexa Fluor 647 Mouse Anti-PTF1A pseudo-colored green and counterstained with DAPI pseudo-colored blue . The nuclei of islet cells DAPI+ stain positively for both NKX2.2 and PTF1A. The nuclei of acinar cells and some islet precursor cells stain positive for PTF1A. All images were analyzed using a BD Pathway 435 High-Content Bioimager System and merged with BD Attovision Software. Show More Panel 1. Two-color flow cytometric analysis of Jurkat cell viability. Jurkat cells were treated with DMSO vehicle Left Plot or 5 muM Camptothecin Right Plot overnight. Cells were resuspended in Annexin V Binding Buffer Cat. No. 556454 and stained with FITC Annexin V Cat. No. 556419 and 0.06 mug/mL DAPI. Camptothecin-treated cells show an increased frequency of apoptotic Annexin V+DAPI- and dead Annexin V+DAPI+ cells. Analysis was performed using a BD LSRFortessa Cell Analyzer System. Panel 2. Flow cytometric analysis of HeLa cell DNA content. Cultured HeLa cells in log phase growth were harvested using Gibco Cell Dissociation Buffer Life Technologies , fixed, and permeabilized using BD Pharmingen Transcription Factor Buffer Set Cat. No. 562574/562725 . Cells were resuspended in DPBS with 1 mug/mL DAPI and analyzed using a low BD LSRFortessa cytometer flow rate. A 405 nm laser with a 450/50 nm bandpass filter was used to collect data; comparable results were obtained using a 355 nm laser with a 450/50 nm bandpass filter not shown . Histograms were deconvoluted by FlowJo software into G0/G1, S, and G2/M populations. Panel 3. Multicolor immunofluorescence analysis of NKX2.2 and PTF1A expression in mouse pancreas. Following antigen retrieval with BD Retrievagen A Buffer Cat. No. 550524 , a formalin-fixed paraffin-embedded pancreas tissue section was stained with Purified Mouse Anti-Nkx2.2 antibody Cat. No. 564731, pseudo-colored red , washed, and stained with Alexa Fluor 488 Goat Anti-Mouse Ig Life Technologies . After washing, the section was stained with Alexa Fluor 647 Mouse Anti-PTF1A pseudo-colored green and counterstained with DAPI pseudo-colored blue . The nuclei of islet cells DAPI+ stain positively for both NKX2.2 and PTF1A. The nuclei of acinar cells and some islet precursor cells stain positive for PTF1A. All images were analyzed using a BD Pathway 435 High-Content Bioimager System and merged with BD Attovision Software.