Preparation And Storage
Recommended Assay Procedures
Flow Cytometry: Mouse BD Fc Block™ purified anti-mouse CD16/CD32 clone 2.4G2 (Cat. No. 553141) may help to reduce non-specific binding to cells bearing Fcγ-receptors. For staining in the presence of Mouse BD Fc Block™, a second step antibody which does not recognize the 2.4G2 mAb (Rat IgG2b, κ) must be used, such as FITC anti-rat IgG2a, mAb RG7/1.30 (Cat. No. 553896).
Caution: Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also effect the results of functional studies, we recommend the NA/LE™ (No Azide/Low Endotoxin) antibody format for in vitro and in vivo use.
Product Notices
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Companion Products
The 53-2.1 monoclonal antibody specifically binds to the CD90.2 (Thy-1.2) alloantigen on thymocytes, most peripheral T lymphocytes, some intraepithelial T lymphocytes (IEL, DEC), epithelial cells, fibroblasts, neurons, hematopoietic stem cells, but not B lymphocytes, of most mouse strains. The 53-2.1 antibody has been reported not to crossreact with Thy-1.1 (e.g., AKR/J, PL), or with rat Thy-1. CD90 is a glycophosphatidylinositol-anchored membrane glycoprotein of the Ig superfamily that is involved in signal transduction. In addition, there is evidence that CD90 mediates adhesion of thymocytes to thymic stroma. The 53-2.1 antibody has been reported to block the binding of the Rat Anti-Mouse CD90.2 antibody (Clone 30-H12) to immobilized thymocyte membranes.
This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
Development References (14)
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Borrello MA, Phipps RP. Differential Thy-1 expression by splenic fibroblasts defines functionally distinct subsets. Cell Immunol. 1996; 173(2):198-206. (Biology). View Reference
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Hasslen SR, Burns AR, Simon SI, et al. Preservation of spatial organization and antigenicity of leukocyte surface molecules by aldehyde fixation: flow cytometry and high-resolution FESEM studies of CD62L, CD11b, and Thy-1. J Histochem Cytochem. 1996; 44(10):1115-1122. (Biology: Electron microscopy). View Reference
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He HT, Naquet P, Caillol D, Pierres M. Thy-1 supports adhesion of mouse thymocytes to thymic epithelial cells through a Ca2(+)-independent mechanism. J Exp Med. 1991; 173(2):515-518. (Biology). View Reference
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Hueber AO, Raposo G, Pierres M, He HT. Thy-1 triggers mouse thymocyte apoptosis through a bcl-2-resistant mechanism. J Exp Med. 1994; 179(3):785-796. (Biology). View Reference
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Ikuta K, Uchida N, Friedman J, Weissman IL. Lymphocyte development from stem cells. Annu Rev Immunol. 1992; 10:759-783. (Biology). View Reference
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Kroczek RA, Gunter KC, Germain RN, Shevach EM. Thy-1 functions as a signal transduction molecule in T lymphocytes and transfected B lymphocytes. Nature. 1986; 322(6075):181-184. (Biology). View Reference
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LeFrancois L. Extrathymic differentiation of intraepithelial lymphocytes: generation of a separate and unequal T-cell repertoire. Immunol Today. 1991; 12(12):436-438. (Biology). View Reference
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Ledbetter JA, Herzenberg LA. Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol Rev. 1979; 47:63-90. (Immunogen: Blocking, Cytotoxicity, Immunoprecipitation). View Reference
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Ledbetter JA, Rouse RV, Micklem HS, Herzenberg LA. T cell subsets defined by expression of Lyt-1,2,3 and Thy-1 antigens. Two-parameter immunofluorescence and cytotoxicity analysis with monoclonal antibodies modifies current views. J Exp Med. 1980; 152(2):280-295. (Biology). View Reference
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Radrizzani M, Carminatti H, Pivetta OH, Idoyaga Vargas VP. Developmental regulation of Thy 1.2 rate of synthesis in the mouse cerebellum. J Neurosci Res. 1995; 42(2):220-227. (Biology). View Reference
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Tigelaar RE, Lewis JM, Bergstresser PR. TCR gamma/delta+ dendritic epidermal T cells as constituents of skin-associated lymphoid tissue. J Invest Dermatol. 1990; 94(6):58S-63S. (Biology). View Reference
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Williams AF, Gagnon J. Neuronal cell Thy-1 glycoprotein: homology with immunoglobulin. Science. 1982; 216(4547):696-703. (Biology). View Reference
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Zheng B, Han S, Kelsoe G. T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1. J Exp Med. 1996; 184(3):1083-1091. (Biology). View Reference
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Zhong RK, Donnenberg AD, Edison L, Harrison DE. The appearance of Thy-1- donor T cells in the peripheral circulation 3-6 weeks after bone marrow transplantation suggests an extrathymic origin. Int Immunol. 1996; 8(2):171-176. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.