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PE BCMA CAR Detection Reagent
572433_2.png

Detection of BCMA CAR expression on transfected CHO cells. CHO cells were transfected with a CAR analog construct, comprised of cell-surface Anti-Human BCMA scFv, an epitope tag, and a transmembrane domain. The cells were then stained with BD™ PE BCMA CAR Detection Reagent (Cat. No. 572803/572433) 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 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.

572433_1.png

Detection of BCMA 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 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™ PE BCMA CAR Detection Reagent (Cat. No. 572803/572433) for 30 minutes at room temperature. DAPI solution (Cat. No. 564907) was added to cells immediately before analysis. Overlapping histograms show BCMA CAR staining with BD™ PE 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.

572433_2.png 572433_1.png

Detection of BCMA CAR expression on transfected CHO cells. CHO cells were transfected with a CAR analog construct, comprised of cell-surface Anti-Human BCMA scFv, an epitope tag, and a transmembrane domain. The cells were then stained with BD™ PE BCMA CAR Detection Reagent (Cat. No. 572803/572433) 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 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. 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™ PE BCMA CAR Detection Reagent (Cat. No. 572803/572433) for 30 minutes at room temperature. DAPI solution (Cat. No. 564907) was added to cells immediately before analysis. Overlapping histograms show BCMA CAR staining with BD™ PE 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.

Product Details
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BD™
Human (QC Testing)
Flow cytometry (Routinely Tested)
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO (GMP)


Preparation And Storage

The recombinant protein was purified from tissue culture supernatant by affinity and size exclusion chromatographies. The recombinant protein was conjugated to the dye under optimum conditions and unconjugated protein and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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Recommended Assay Procedures

Surface Staining Protocol

• Resuspend up to 1 × 10^6 cells in 100 µl of stain buffer.

• Add 5 µl of the BCMA CAR Detection Reagent and incubate at either 4°C or at RT for 30 mins protected from light.

• 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).

Bead-based compensation or unmixing controls, such as BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies or IgG Fc-fusion proteins. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.

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Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. 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.
  4. 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).
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. For U.S. patents that may apply, see bd.com/patents.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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572433 Rev. 3
Antibody Details
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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 with the functional capabilities 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 proliferate 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. BCMA CAR Detection Reagent is a recombinant Human BCMA (aa 1-54, extracellular domain)-Mouse IgG2a Fc fusion protein conjugated to a fluorochrome that enables 1-step staining, alone or in multicolor panels, for flow cytometric analysis of BMCA CAR-expressing T, NK, or other cells.

572433 Rev. 3
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
496 nm, 566 nm
576 nm
572433 Rev.3
Citations & References
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View product citations for antibody "572433" on CiteAb

Development References (5)

  1. Ali SA, Shi V, Maric I, et al. T cells expressing an anti-B-cell maturation antigen chimeric antigen receptor cause remissions of multiple myeloma.. Blood. 2016; 128(13):1688-700. (Biology). View Reference
  2. Hansen DK, Peres LC, Dima D, et al. Comparison of Standard-of-Care Idecabtagene Vicleucel and Ciltacabtagene Autoleucel in Relapsed/Refractory Multiple Myeloma.. J Clin Oncol. 2025; 43(13):1597-1609. (Biology). View Reference
  3. Hu Y, Huang J. The Chimeric Antigen Receptor Detection Toolkit.. Front Immunol. 2020; 11:1770. (Biology). View Reference
  4. Scheller L, Tebuka E, Rambau PF, et al. BCMA CAR-T cells in multiple myeloma-ready for take-off?. Leuk Lymphoma. 2024; 65(2):143-157. (Biology). View Reference
  5. Suarez ER, Chang de K, Sun J, et al. Chimeric antigen receptor T cells secreting anti-PD-L1 antibodies more effectively regress renal cell carcinoma in a humanized mouse model.. Oncotarget. 2016; 7(23):34341-55. (Biology). View Reference
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572433 Rev. 3

Please refer to Support Documents for Quality Certificates

 

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

 

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures. 

 

Although not required, these products are manufactured in accordance with Good Manufacturing Practices.