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BUV661 Biosimilar Anti-Human CD52

BD OptiBuild™ BUV661 Biosimilar Anti-Human CD52

Clone Alemtuzumab297.rMAb (also known as Alemtuzumab N297A Biosimilar.rMAb)

Product Details
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BD OptiBuild™
CAMPATH-1 antigen; HE5
Human (Tested in Development)
Human IgG1, κ
Human CD52
Flow cytometry (Qualified)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.

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 that minimize unconjugated dye and antibody. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

Product Notices

  1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  2. Please refer to for technical protocols.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at
  7. 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
  8. Please refer to to access safety data sheets (SDS).
  9. For U.S. patents that may apply, see
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Antibody Details
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The Alemtuzumab297.rMAb  (also known as, Alemtuzumab N297A Biosimilar.rMAb) is a research grade humanized recombinant human IgG1, kappa antibody that specifically recognizes the extracellular domain of human CD52, similarly to the therapeutic Alemtuzumab antibody. The Alemtuzumab297.rMAb uses variable region sequences from Alemtuzumab and constant region consensus sequences from 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.  The CD52 antigen is a 25-29 kDa N-glycosylated, GPI-linked protein that is also known as Cambridge pathology antigen 1 (CAMPATH-1 antigen), or Human epididymis-specific protein 5 (He5).  It is expressed on thymocytes, lymphocytes, monocytes, macrophages, dendritic cells, and eosinophils, but is weakly or not expressed on neutrophils, erythrocytes, platelets, or hematopoietic stem cells. CD52 is variably expressed on malignant lymphoid cells and serves as a target for therapeutic CD52-specific antibodies.

The Alemtuzumab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.

758156 Rev. 1
Format Details
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The BD Horizon Brilliant™ Ultraviolet 661 (BUV661) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 660-nm. BUV661, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 660-nm (e.g., 670/25 bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Ultraviolet 355 nm
350 nm
660 nm
758156 Rev.1
Citations & References
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Development References (10)

  1. Bologna L, Gotti E, Manganini M, et al. Mechanism of action of type II, glycoengineered, anti-CD20 monoclonal antibody GA101 in B-chronic lymphocytic leukemia whole blood assays in comparison with rituximab and alemtuzumab.. J Immunol. 2011; 186(6):3762-9. (Biology). View Reference
  2. Golay J, Manganini M, Rambaldi A, Introna M. Effect of alemtuzumab on neoplastic B cells.. Haematologica. 2004; 89(12):1476-83. (Biology). View Reference
  3. Hale G. Synthetic peptide mimotope of the CAMPATH-1 (CD52) antigen, a small glycosylphosphatidylinositol-anchored glycoprotein.. Immunotechnology. 1995; 1(3-4):175-87. (Biology). View Reference
  4. Havari E, Turner MJ, Campos-Rivera J, et al. Impact of alemtuzumab treatment on the survival and function of human regulatory T cells in vitro.. Immunology. 2014; 141(1):123-31. (Biology). View Reference
  5. Lowenstein H, Shah A, Chant A, Khan A. Different mechanisms of Campath-1H-mediated depletion for CD4 and CD8 T cells in peripheral blood.. Transpl Int. 2006; 19(11):927-36. (Biology). View Reference
  6. Mohan SR, Clemente MJ, Afable M, et al. Therapeutic implications of variable expression of CD52 on clonal cytotoxic T cells in CD8+ large granular lymphocyte leukemia.. Haematologica. 2009; 94(10):1407-14. (Biology). View Reference
  7. Pernick N. CD50-59. 2023. Available: 10/162023.
  8. Riechmann V, van Crüchten I, Sablitzky F. The expression pattern of Id4, a novel dominant negative helix-loop-helix protein, is distinct from Id1, Id2 and Id3. Nucleic Acids Res. 1994; 22(5):749-755. (Biology). View Reference
  9. Toh BH, Kyaw T, Tipping P, Bobik A. Immune regulation by CD52-expressing CD4 T cells.. Cell Mol Immunol. 2013; 10(5):379-82. (Biology). View Reference
  10. Trzonkowski P, Zilvetti M, Friend P, Wood KJ. Recipient memory-like lymphocytes remain unresponsive to graft antigens after CAMPATH-1H induction with reduced maintenance immunosuppression.. Transplantation. 2006; 82(10):1342-51. (Biology). View Reference
View All (10) View Less
758156 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.