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BUV496 Rat Anti-Mouse CD45
Product Details
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BD OptiBuild™
Cd45; L-CA; LCA; Ly-5; Ly5; Ptprc
Mouse (Tested in Development)
Rat LEW, also known as Lewis IgG2b, λ
Mouse Lymphoma Cell Line
Flow cytometry (Qualified)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.

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 CompBead 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 two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  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. Researchers should determine the optimal concentration of this reagent for their individual applications.
  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
  6. Please refer to for technical protocols.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to to access safety data sheets (SDS).
  9. BD Horizon Brilliant Ultraviolet 496 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
752411 Rev. 1
Antibody Details
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The mAb I3/2.3 recognizes mouse CD45, which is also known as the leukocyte common antigen (L-CA or LCA), T200, or Lymphocyte antigen 5 (Ly-5). CD45 is a 180-240 kDa single-pass type I transmembrane glycoprotein that is encoded by Ptprc (protein tyrosine phosphatase, receptor type, C) which belongs to the protein tyrosine phosphatase (PTP) family. CD45 is variably expressed on thymocytes and other hematopoietic cells except mature erythrocytes and platelets. Different isoforms of CD45 arise from alternative splicing of variable exons 4, 5, and 6, which encode A, B, and C determinants, respectively, that encode extracellular domains of the transmembrane receptor. The differential expression of these isoforms is related to the cell's lineage as well as its state of activation and maturation. The mAb I3/2.3 reportedly recognizes all CD45 isoforms. CD45 plays key roles as a protein tyrosine-protein phosphatase in leucocyte responses including signal pathways transduced by antigen specific receptors expressed on T cells (TCRs) and B cells (BCRs).

The antibody was conjugated to BD Horizon™ BUV496 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 496-nm. BD Horizon BUV496 can be excited by the ultraviolet laser (355 nm) and detected with a 515/30 nm filter with a 450LP. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into the channel detecting BD Horizon V500 or BV510 (eg, 525/40-nm filter). However, the spillover can be corrected through compensation as with any other dye combination.

752411 Rev. 1
Format Details
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The BD Horizon Brilliant™ Ultraviolet 496 (BUV496) 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 496-nm. BUV496, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 500-nm (e.g., 515/30-nm bandpass filter). The acceptor dye can be excited by the Violet (405-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
496 nm
752411 Rev.1
Citations & References
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Development References (5)

  1. Alaverdi N. Monoclonal antibodies to mouse cell-surface antigens.. Curr Protoc Immunol. 2002; Appendix 4:Appendix 4B. (Clone-specific). View Reference
  2. Johnson P, Greenbaum L, Bottomly K, Trowbridge IS. Identification of the alternatively spliced exons of murine CD45 (T200) required for reactivity with B220 and other T200-restricted antibodies. J Exp Med. 1989; 169(3):1179-1184. (Clone-specific: Flow cytometry). View Reference
  3. Lefrancois L, Goodman T. Developmental sequence of T200 antigen modifications in murine T cells.. J Immunol. 1987; 139(11):3718-24. (Clone-specific: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
  4. Takedachi M, Qu D, Ebisuno Y, et al. CD73-generated adenosine restricts lymphocyte migration into draining lymph nodes.. J Immunol. 2008; 180(9):6288-96. (Biology). View Reference
  5. Trowbridge IS. Interspecies spleen-myeloma hybrid producing monoclonal antibodies against mouse lymphocyte surface glycoprotein, T200.. J Exp Med. 1978; 148(1):313-23. (Immunogen: Immunoprecipitation, Ribonuclease protection assay). View Reference
View All (5) View Less
752411 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.