BD™ Alexa Fluor™ 488 CD19 CAR Detection Reagent
Detection of CD19 CAR expression on transfected CHO cells. CHO cells were transfected with a CAR analog construct, comprised of cell-surface Anti-Human CD19 scFv, an epitope tag, and a transmembrane domain. The cells were then stained with BD™ Alexa Fluor™ 488 CD19 CAR Detection Reagent (Cat. No. 572813/572814) and Alexa Fluor™ 647 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 CD19 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 CD19 CAR expression on transduced primary Human T cells. 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 CD19 CAR lentivirus, expanded for 7-10 days in culture with IL-2, and cryopreserved. Thawed CD19 CAR-transduced cells or mock transduced cells were stained with BD™ Alexa Fluor™ 488 CD19 CAR Detection Reagent (Cat. No. 572813/572814) for 30 minutes at room temperature. DAPI solution (Cat. No. 564907) was added to cells immediately before analysis. Overlapping histograms show CD19 CAR staining with BD™ Alexa Fluor 488TM CD19 CAR Detection Reagent of either CD19 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 CD19 CAR expression on transfected CHO cells. CHO cells were transfected with a CAR analog construct, comprised of cell-surface Anti-Human CD19 scFv, an epitope tag, and a transmembrane domain. The cells were then stained with BD™ Alexa Fluor™ 488 CD19 CAR Detection Reagent (Cat. No. 572813/572814) and Alexa Fluor™ 647 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 CD19 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 CD19 CAR expression on transduced primary Human T cells. 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 CD19 CAR lentivirus, expanded for 7-10 days in culture with IL-2, and cryopreserved. Thawed CD19 CAR-transduced cells or mock transduced cells were stained with BD™ Alexa Fluor™ 488 CD19 CAR Detection Reagent (Cat. No. 572813/572814) for 30 minutes at room temperature. DAPI solution (Cat. No. 564907) was added to cells immediately before analysis. Overlapping histograms show CD19 CAR staining with BD™ Alexa Fluor 488TM CD19 CAR Detection Reagent of either CD19 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.
Preparation And Storage
Recommended Assay Procedures
Surface Staining Protocol
• Aliquot 100 μl of human whole blood into tubes, or up to 1 × 10^6 nucleated cells (including human PBMC or other cell samples such as CD19 CAR-expressing cells) per 100 μl of stain buffer into microwell plates or tubes for staining.
• Add 5 μl of the CD19 CAR Detection Reagent and incubate at either 4°C or at RT for 30 mins protected from light.
• For stained whole blood samples, treat with either BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) or with BD FACS™ Lysing Solution (Cat. No. 349202). Wash cells with stain buffer and proceed with acquisition.
• For stained PBMC or other nucleated cell samples, wash cells with stain buffer, add viability dye and proceed with acquisition.
Notes:
• Recommend using BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564220) prior to staining as needed.
• This CAR Detection Reagent has been tested for compatibility with either BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ Buffer (Cat. No. 554723) or the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574).
• Avoid co-stains with Human CD19- or CD81-specific antibodies when using this reagent.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- For U.S. patents that may apply, see bd.com/patents.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
Companion Products
Chimeric Antigen Receptors (CAR) are engineered transmembrane proteins containing an extracellular single-chain domain from a proven antibody fused with major intracellular signaling components from the T Cell Receptor (TCR) complex. A CAR thus combines a defined antibody specificity and affinity with the cellular functionality of an effector cell and is a popular approach for cellular immunotherapy of cancer or autoimmunity. In CAR T cell therapy, T cells are isolated from a patient's blood, activated, transduced with a CAR gene, expanded, and reinfused into the patient, where they expand upon encounter with target antigen and eliminate antigen-expressing cells. Flow cytometry is used throughout this workflow to assess CAR expression and CAR-T phenotype pre-infusion, and post-infusion to probe expansion, persistence, and in vivo CAR-T characteristics. CD19 CAR Detection Reagent is a recombinant Human CD19 (aa 21-278, extracellular domain)-CD81 (LEL) - human IgG1 (N297A) Fc fusion protein conjugated to a fluorochrome that enables 1-step staining, alone or in multicolor panels, for flow cytometric analysis of CD19 CAR-expressing T, NK, or other cells.
Development References (7)
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De Oliveira SN, Wang J, Ryan C, Morrison SL, Kohn DB, Hollis RP. A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors.. J Transl Med. 2013; 11:23. (Biology). View Reference
-
Faramand RG, Locke FL. Intermediate-affinity CD19-directed CAR T cell product obecabtagene autoleucel demonstrates favourable safety and efficacy in R/R B-ALL.. Nat Rev Clin Oncol. 2025; 22(3):161-162. (Biology). View Reference
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Hu Y, Huang J. The Chimeric Antigen Receptor Detection Toolkit.. Front Immunol. 2020; 11:1770. (Biology). View Reference
-
Ma S, Li X, Wang X, et al. Current Progress in CAR-T Cell Therapy for Solid Tumors.. Int J Biol Sci. 2019; 15(12):2548-2560. (Biology). View Reference
-
Patel KK, Tariveranmoshabad M, Kadu S, Shobaki N, June C. From concept to cure: The evolution of CAR-T cell therapy.. Mol Ther. 2025; 33(5):2123-2140. (Biology). View Reference
-
Rampotas A, Richter J, Isenberg D, Roddie C. CAR-T cell therapy embarks on autoimmune disease.. Bone Marrow Transplant. 2025; 60(1):6-9. (Biology). View Reference
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Yuan X, Wang F, Zhao P, et al. Efficacy and safety of CD19 combined with CD22 or CD20 chimeric antigen receptor T-cell therapy for hematological malignancies.. Front Immunol. 2025; 16:1577360. (Biology). View Reference
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