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BUV615 Mouse Anti-Mouse CD72 a, b, and d Alloantigens
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Product Details
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BD OptiBuild™
Lyb-2.1; Lyb-2.2; and Lyb-2.4
Mouse (Tested in Development)
Mouse SJL IgG2b, κ
(C57BL/6 • DBA/2)F1 hybrid mouse pre-B-cell leukemia line 70Z/3
Flow cytometry (Qualified)
0.2 mg/ml
12517
AB_2875884
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


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 www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources 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 http://regdocs.bd.com to access safety data sheets (SDS).
  9. CF™ is a trademark of Biotium, Inc.
  10. BD Horizon Brilliant Ultraviolet 615 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.
752367 Rev. 1
Antibody Details
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K10.6

The K10.6 monoclonal antibody specifically recognizes Lyb-2.1, Lyb-2.2, and Lyb-2.4 (CD72 a, b, and d alloantigens, respectively). CD72 is a 45-kDa type-II membrane protein, containing a C-type lectin-like domain and ITIM and ITIM-like sequences in the cytoplasmic tail. CD72 is expressed at all stages of B-lymphocyte development except plasma cells, and it has been shown to negatively regulate B-cell receptor signaling. Analysis of CD72-deficient mice supports these results and shows that CD72 is involved in B-cell development. CD72-stimulated B cells show a transient association of CD19 with CD72, as well as an increase in Tyr-phosphorylation of CD19. CD72 is reported to be a ligand for CD5, although this is controversial. It is also reported to be a ligand for CD100 (Sema4D). The CD72 alloantigens Lyb-2.1 (originally identified as Ly-m19.2), Lyb-2.2 (originally identified as Ly-32.2), Lyb-2.3, and Lyb-2.4 are encoded by the Cd72[a], Cd72[b], Cd72[c], and Cd72[d] alleles, respectively. Lyb-2.1 is expressed on B lymphocytes of CBA/J, C3H/Bi, C57BR, C57L, C58, DBA/1, DBA/2, and SWR strains. Lyb-2.2 is expressed on B lymphocytes, a subset of peripheral T cells, and activated T lymphocytes in A, BALB/c, CBA/H, C3H/He, C57BL, PL, and 129 strains. Lyb-2.4 is expressed on a subset of splenocytes of the STS/A strain. K10.6 antibody does not react with Lyb-2.3 (AKR and SJL strains) nor with non-lymphoid tissues. Five serological specificities of CD72 alloantigens have been described and the nomenclature CD72.1, CD72.2, CD72.3, CD72.4, and CD72.5 proposed, which does not correspond to the names of the alleles. Some authors have referred to the specificity of mAb K10.6 as CD72.4.

The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP.  Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).

752367 Rev. 1
Format Details
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BUV615
The BD Horizon Brilliant™ Ultraviolet 615 (BUV615) 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 615-nm. BUV615, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 615-nm (e.g, 610/20 bandpass filter). The acceptor dye can be excited by the Blue (488-nm) and yellow-green (561-nm) lasers resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV615
Ultraviolet 355 nm
350 nm
615 nm
752367 Rev.1
Citations & References
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Development References (12)

  1. Adachi T, Wakabayashi C, Nakayama T, Yakura H, Tsubata T. CD72 negatively regulates signaling through the antigen receptor of B cells.. J Immunol. 2000; 164(3):1223-9. (Biology). View Reference
  2. Bikah G, Lynd FM, Aruffo AA, Ledbetter JA, Bondada S. A role for CD5 in cognate interactions between T cells and B cells, and identification of a novel ligand for CD5. Int Immunol. 1998; 10(8):1185-1196. (Biology). View Reference
  3. Doolittle, D.P., M.T. Davisson, J.N. Guidi, and M.C. Green. M.F. Lyon, S. Rastan and S.D.M. Brown, ed. Genetic variants and strains of the laboratory mouse. Oxford : Oxford University Press; 1996:133.
  4. Kumanogoh A, Watanabe C, Lee I, et al. Identification of CD72 as a lymphocyte receptor for the class IV semaphorin CD100: a novel mechanism for regulating B cell signaling. Immunity. 2000; 13(5):621-631. (Biology). View Reference
  5. Luo W, Van de Velde H, von Hoegen I, Parnes JR, Thielemans K. Ly-1 (CD5), a membrane glycoprotein of mouse T lymphocytes and a subset of B cells, is a natural ligand of the B cell surface protein Lyb-2 (CD72). J Immunol. 1992; 148(6):1630-1634. (Biology). View Reference
  6. Pan C, Baumgarth N, Parnes JR. CD72-deficient mice reveal nonredundant roles of CD72 in B cell development and activation. Immunity. 1999 October; 11(4):495-506. (Biology). View Reference
  7. Robinson WH, Landolfi MM, Parnes JR. Allele-specific expression of the mouse B-cell surface protein CD72 on T cells. Immunogenetics. 1997; 75(3):195-200. (Biology). View Reference
  8. Robinson WH, Landolfi MM, Schäfer H, Parnes JR. Biochemical identity of the mouse Ly-19.2 and Ly-32.2 alloantigens with the B cell differentiation antigen Lyb-2/CD72. J Immunol. 1993 November; 151(9):4764-4772. (Biology). View Reference
  9. Robinson WH, Ying H, Miceli MC, Parnes JR. Extensive polymorphism in the extracellular domain of the mouse B cell differentiation antigen Lyb-2/CD72.. J Immunol. 1992; 149(3):880-6. (Biology). View Reference
  10. Tada N, Kimura S, Liu Y, Taylor BA, Hämmerling U. Ly-m19: the Lyb-2 region of mouse chromosome 4 controls a new surface alloantigen. Immunogenetics. 1981; 13(6):539-546. (Immunogen). View Reference
  11. Venkataraman C, Lu PJ, Buhl AM, Chen CS, Cambier JC, Bondada S. CD72-mediated B cell activation involves recruitment of CD19 and activation of phosphatidylinositol 3-kinase. Eur J Immunol. 1998 October; 28(10):3003-3016. (Biology). View Reference
  12. Ying H, Healy JI, Goodnow CC, Parnes JR. Regulation of mouse CD72 gene expression during B lymphocyte development. J Immunol. 1998; 161(9):4760-4767. (Biology). View Reference
View All (12) View Less
752367 Rev. 1

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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.