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      BUV395 Biosimilar Anti-Human CD20 Rituximab297

      BD Horizon™ BUV395 Biosimilar Anti-Human CD20 Rituximab297

      Clone Rituximab297.rMAb (also known as Rituximab N297A Biosimilar.rMAb)

      (RUO)
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      BUV395 Biosimilar Anti-Human CD20 Rituximab297
      Two-color flow cytometric analysis of Human CD20 (Rituximab Biosimilar) expression on Human peripheral blood lymphocytes.  Human whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415) and with either BD OptiBuild™ BUV395 IgG1, κ N297A Human Isotype Control (Cat. No. 756237; Left Plot) or BD Horizon™ BUV395 Biosimilar Anti-Human CD20 Rituximab297 antibody (Cat. No. 570360/570430; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD20 (Rituximab Biosimilar) [or Ig Isotype control staining] versus CD19 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      BUV395 Biosimilar Anti-Human CD20 Rituximab297
      Multiparameter flow cytometric analysis of Human CD20 (Rituximab Biosimilar) expression on Human peripheral blood leukocytes.  Human whole blood was stained with either BD OptiBuild™ BUV395 IgG1, κ N297A Human Isotype Control (Cat. No. 756237; Left Plot) or BD Horizon™ BUV395 Biosimilar Anti-Human CD20 Rituximab297 antibody (Cat. No. 570360/570430; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD20 (Rituximab Biosimilar) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      BUV395 Biosimilar Anti-Human CD20 Rituximab297 BUV395 Biosimilar Anti-Human CD20 Rituximab297
      Two-color flow cytometric analysis of Human CD20 (Rituximab Biosimilar) expression on Human peripheral blood lymphocytes.  Human whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415) and with either BD OptiBuild™ BUV395 IgG1, κ N297A Human Isotype Control (Cat. No. 756237; Left Plot) or BD Horizon™ BUV395 Biosimilar Anti-Human CD20 Rituximab297 antibody (Cat. No. 570360/570430; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD20 (Rituximab Biosimilar) [or Ig Isotype control staining] versus CD19 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Multiparameter flow cytometric analysis of Human CD20 (Rituximab Biosimilar) expression on Human peripheral blood leukocytes.  Human whole blood was stained with either BD OptiBuild™ BUV395 IgG1, κ N297A Human Isotype Control (Cat. No. 756237; Left Plot) or BD Horizon™ BUV395 Biosimilar Anti-Human CD20 Rituximab297 antibody (Cat. No. 570360/570430; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD20 (Rituximab Biosimilar) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leukocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Product Details
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      BD Horizon™
      B1; Bp35; LEU-16; MS4A1
      Human (QC Testing)
      Human IgG1, κ
      Human CD20
      Flow cytometry (Routinely Tested)
      5 µl/test
      931
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody 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

        BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

        For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet for BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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).
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. 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.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
      7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      8. For U.S. patents that may apply, see bd.com/patents.
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      570360 Rev. 1
      Antibody Details
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      Rituximab297.rMAb

      The Rituximab297.rMAb (also known as, Rituximab N297A Biosimilar.rMAb) is a research grade chimeric recombinant human IgG1, kappa antibody that specifically recognizes the extracellular domain of human CD20 similarly to the therapeutic Rituximab antibody. The Rituximab297.rMAb uses the same variable region sequences as Rituximab, combined with constant region sequences derived from consensus sequences of human IgG1, kappa. The asparagine at position 297 of the constant heavy chain has been replaced with alanine (N297A) to further reduce Fc receptor interactions of this reagent. Rituximab triggers target cell death through multiple mechanisms including complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and apoptosis. The therapeutic Rituximab has been approved to treat human B cell malignancies, autoimmune disorders, and transplant rejection. CD20 is a 33-37 kDa, unglycosylated four-transmembrane phosphoprotein with a cytoplasmic N-terminus and C-terminus that is encoded by MS4A1 (Membrane-spanning 4-domains, subfamily A, member 1). CD20 is expressed on pre-B-cells, naive and activated B cells, memory B cells, and at lower levels on a small subset of T cells but is absent from plasmablasts and plasma cells. It is also expressed on most malignant B cells and CD20-positive T-cell lymphomas. CD20 functions in mediating calcium transport and B cell activation, differentiation, and survival.

         The Rituximab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.

      570360 Rev. 1
      Format Details
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      BUV395
      The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BUV395
      Ultraviolet 355 nm
      348 nm
      395 nm
      570360 Rev.1
      Citations & References
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      View product citations for antibody "570360" on CiteAb

      Development References (10)

      1. Almasri NM, Duque RE, Iturraspe J, Everett E, Braylan RC. Reduced expression of CD20 antigen as a characteristic marker for chronic lymphocytic leukemia. Am J Hematol. 1992; 40:259-263. (Biology).
      2. Brekke OH, Sandlie I. Therapeutic antibodies for human diseases at the dawn of the twenty-first century.. Nat Rev Drug Discov. 2003; 2(1):52-62. (Biology). View Reference
      3. Cragg MS, Walshe CA, Ivanov AO, Glennie MJ. The biology of CD20 and its potential as a target for mAb therapy. Curr Dir Autoimmun. 2005; 8:140-174. (Biology). View Reference
      4. Deans JP, Li H, Polyak MJ. CD20-mediated apoptosis: signalling through lipid rafts. Immunology. 2002; 107:176-182. (Biology).
      5. Golay J, Zaffaroni L, Vaccari T, et al. Biologic response of B lymphoma cells to anti-CD20 monoclonal antibody rituximab in vitro: CD55 and CD59 regulate complement-mediated cell lysis. Blood. 2000; 95:3900-3908. (Biology).
      6. Hultin LE, Hausner MA, Hultin PM, Giorgi JV. CD20 (pan-B cell) antigen is expressed at a low level on asubpopulation of human T lymphocytes. Cytometry. 1993; 14:196-204. (Biology).
      7. Kennedy AD, Solga MD, Schuman TA, et al. An anti-C3b(i) mAb enhances complement activation, C3b(i) deposition, and killing of CD20+ cells by rituximab.. Blood. 2003; 101(3):1071-9. (Biology). View Reference
      8. Kitamura A, Yamashita Y, Mori N. CD20-positive cytotoxic T cell lymphoma: report of two cases and review of the literature. J Clin Exp Hematop. 2005; 45(1):45-50. (Biology).
      9. Uchida J, Hamaguchi Y, Oliver JA, et al. The innate mononuclear phagocyte network depletes B lymphocytes through Fc receptor-dependent mechanisms during anti-CD20 antibody immunotherapy.. J Exp Med. 2004; 199(12):1659-1669. (Biology). View Reference
      10. Zola H, Swart B, Nicholson I, Voss E. CD20. In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:72.
      View All (10) View Less
      570360 Rev. 1

      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.