BDTM Alexa FluorTM 647 BCMA CAR Detection Reagent

Description

Detection of BCMA CAR expression on transfected CHO cells. CHO cells were transfected with a CAR analog construct, comprised of cell-surface expressed Anti-Human BCMA scFv, an epitope tag, and a transmembrane domain. The cells were then stained with BD Alexa Fluor 647 BCMA CAR Detection Reagent Cat. No. 572805/572616 and BD OptiBuild RY586 epitope tag-specific antibody for 30 minutes at room temperature. DAPI Solution Cat. No. 564907 was added to cells immediately prior to analysis. The bivariate pseudocolor density plot shows BCMA CAR expression correlated with epitope tag-specific staining, gated on viable DAPI-negative CHO cells Flow cytometry was performed using a BD LSRFortessa Cell Analyzer System and FlowJo Software. Detection of BCMA CAR expression on transduced primary Human T cells. Human T cells from a healthy donor were purified using the BD IMag Human T Lymphocyte Enrichment Set-DM Cat. No. 557874 . T cells were then activated with Anti-CD3/CD28 antibody-coupled beads plus purified BD Pharmingen Recombinant Human IL-2 protein Cat. No. 554603 for 2 days, infected with BCMA CAR lentivirus, expanded for 7-10 days in culture with IL-2, and cryopreserved. Thawed BCMA CAR-transduced cells or mock transduced cells were stained with BD Alexa Fluor 647 BCMA CAR Detection Reagent Cat. No. 572805/572616 for 30 minutes at room temperature. DAPI solution Cat. No. 564907 was added to cells immediately prior to analysis. Overlapping histograms show BCMA CAR staining with BD Alexa Fluor 647 BCMA CAR Detection Reagent of either BCMA CAR-transduced cells solid line versus mock transduced cells dashed line , gated on viable DAPI-negative cells. Flow cytometry was performed using a BD LSRFortessa Cell Analyzer System and FlowJo Software. Detection of BCMA CAR expression on transfected CHO cells. CHO cells were transfected with a CAR analog construct, comprised of cell-surface expressed Anti-Human BCMA scFv, an epitope tag, and a transmembrane domain. The cells were then stained with BD Alexa Fluor 647 BCMA CAR Detection Reagent Cat. No. 572805/572616 and BD OptiBuild RY586 epitope tag-specific antibody for 30 minutes at room temperature. DAPI Solution Cat. No. 564907 was added to cells immediately prior to analysis. The bivariate pseudocolor density plot shows BCMA CAR expression correlated with epitope tag-specific staining, gated on viable DAPI-negative CHO cells Flow cytometry was performed using a BD LSRFortessa Cell Analyzer System and FlowJo Software. Show More Detection of BCMA CAR expression on transduced primary Human T cells. Human T cells from a healthy donor were purified using the BD IMag Human T Lymphocyte Enrichment Set-DM Cat. No. 557874 . T cells were then activated with Anti-CD3/CD28 antibody-coupled beads plus purified BD Pharmingen Recombinant Human IL-2 protein Cat. No. 554603 for 2 days, infected with BCMA CAR lentivirus, expanded for 7-10 days in culture with IL-2, and cryopreserved. Thawed BCMA CAR-transduced cells or mock transduced cells were stained with BD Alexa Fluor 647 BCMA CAR Detection Reagent Cat. No. 572805/572616 for 30 minutes at room temperature. DAPI solution Cat. No. 564907 was added to cells immediately prior to analysis. Overlapping histograms show BCMA CAR staining with BD Alexa Fluor 647 BCMA CAR Detection Reagent of either BCMA CAR-transduced cells solid line versus mock transduced cells dashed line , gated on viable DAPI-negative cells. Flow cytometry was performed using a BD LSRFortessa Cell Analyzer System and FlowJo Software. Show More Detection of BCMA CAR expression on transfected CHO cells. CHO cells were transfected with a CAR analog construct, comprised of cell-surface expressed Anti-Human BCMA scFv, an epitope tag, and a transmembrane domain. The cells were then stained with BD Alexa Fluor 647 BCMA CAR Detection Reagent Cat. No. 572805/572616 and BD OptiBuild RY586 epitope tag-specific antibody for 30 minutes at room temperature. DAPI Solution Cat. No. 564907 was added to cells immediately prior to analysis. The bivariate pseudocolor density plot shows BCMA CAR expression correlated with epitope tag-specific staining, gated on viable DAPI-negative CHO cells Flow cytometry was performed using a BD LSRFortessa Cell Analyzer System and FlowJo Software. Detection of BCMA CAR expression on transduced primary Human T cells. Human T cells from a healthy donor were purified using the BD IMag Human T Lymphocyte Enrichment Set-DM Cat. No. 557874 . T cells were then activated with Anti-CD3/CD28 antibody-coupled beads plus purified BD Pharmingen Recombinant Human IL-2 protein Cat. No. 554603 for 2 days, infected with BCMA CAR lentivirus, expanded for 7-10 days in culture with IL-2, and cryopreserved. Thawed BCMA CAR-transduced cells or mock transduced cells were stained with BD Alexa Fluor 647 BCMA CAR Detection Reagent Cat. No. 572805/572616 for 30 minutes at room temperature. DAPI solution Cat. No. 564907 was added to cells immediately prior to analysis. Overlapping histograms show BCMA CAR staining with BD Alexa Fluor 647 BCMA CAR Detection Reagent of either BCMA CAR-transduced cells solid line versus mock transduced cells dashed line , gated on viable DAPI-negative cells. Flow cytometry was performed using a BD LSRFortessa Cell Analyzer System and FlowJo Software.