Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) 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).
Product Notices
- This antibody was developed for use in flow cytometry.
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
Companion Products
The JY/93 monoclonal antibody specifically recognizes Lyb-2.2 and Lyb-2.3 (CD72 b and c alloantigens, respectively). CD72 is a 45-kDa type-II membrane protein containing a C-type lectin-like domain, an ITIM and ITIM-like sequence in the cytoplasmic tail. Recently, CD72 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 CD5,5 although this is controversial. 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-2.2 (CD72b) is expressed on B lymphocytes, a subset of peripheral T cells, activated thymocytes, and activated T lymphocytes in A, BALB/c, CBA/H, CBA/N, C3H/He, C57BL, NZB, NZW, PL, and 129 strains. Moreover, expression of CD72b on T cells is dominant in F1 hybrid mice, and its level of expression increases upon activation. Lyb-2.3 (CD72c) is expressed on B cells of AKR, CE, SJL, and other mouse strains. Although staining of a small percentage of splenic T cells has been detected in CD72c mice, northern blot analysis has shown no expression of CD72c in T lymphocytes of these mice. JY/93 mAb does not stain cells from CD72a mice (e.g., CBA/J, DBA/2, SWR), and it does not stain resting thymocytes, splenic CD11b-positive cells, or NK cells. Five serological specificities of CD72 alloantigens have been described and the nomenclature CD72.1, CD72.2, CD72.3, CD72.4, and CD72.5 proposed.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (10)
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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
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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
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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.
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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
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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
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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
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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
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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
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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
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Yamashita Y, Phee H, Tudor KS, et al. A unique CD72 epitope suggests a potential interaction with Fc gamma RII/CD32 on B lineage lymphocytes.. Hybridoma (Larchmt). 2006; 25(3):107-14. (Immunogen: Flow cytometry, Functional assay, Immunofluorescence, Immunoprecipitation). View Reference
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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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.