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 Ultraviolet 737 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.
Companion Products
The D12 monoclonal antibody specifically binds to CD11b which is also known as Integrin alpha M (Integrin αM), Mac-1 subunit alpha (Mac-1a) or complement receptor 3 alpha chain (CR3a). CD11b is encoded by ITGAM (integrin subunit alpha M) and belongs to the integrin alpha subunit gene family. CD11b is expressed as a ~165-kDa type I transmembrane glycoprotein that associates with the ~95-kDa integrin β2 (CD18) to form the heterodimeric CD11b/CD18 (αM/β2) complex which also known as Mac-1 or CR3. CD11b is expressed on monocytes, dendritic cells, granulocytes, as well as on some T cells, B cells, and NK cells. CD11b functions in cell-cell and cell-substrate interactions and is a receptor for multiple ligands including inactivated C3b (iC3b), ICAM-1 (CD54), ICAM-2 (CD102), ICAM-3 (CD50) or fibrinogen. Mac-1 regulates leucocyte adhesion and migration, as well as phagocytosis of opsonized particles.
CAUTION Binding of this CD11b antibody depends on the presence of Ca++. EDTA or ACD, as anticoagulant might affect binding. Using heparin as an anticoagulant or removal of the anticoagulant is recommended.
The antibody was conjugated to BD Horizon™ BUV737 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 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
Development References (10)
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Lanier LL, Phillips JH. A map of the cell surface antigens expressed on resting and activated human natural killer cells. In: Reinherz EL. Ellis L. Reinherz .. et al., ed. Leukocyte typing II. New York: Springer-Verlag; 1986:157-170.
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Bernstein ID, Self S. Joint report of the Myeloid Section of the Second International Workshop on Human Leukocyte Differentiation Antigens. In: Reinherz EL, Haynes BF, Nadler LM, Bernstein ID, ed. Leukocyte Typing II: Human Myeloid and Hematopoietic Cells. New York, NY: Springer-Verlag; 1986:1-25.
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Bray RA, Gottschalk LR, Landay AL, Gebel HM. Differential surface marker expression in patients with CD-16+ lymphoproliferative disorders: in vivo model for NK differentiation.. Hum Immunol. 1987; 19(2):105-15. (Biology). View Reference
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Clement LT, Grossi CE, Gartland GL. Morphologic and phenotypic features of the subpopulation of Leu-2+ cells that suppresses B cell differentiation.. J Immunol. 1984; 133(5):2461-8. (Clone-specific). View Reference
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Gebel HM, Kaizer H, Landay AL. Characterization of circulating suppressor T lymphocytes in bone marrow transplant recipients.. Transplantation. 1987; 43(2):258-63. (Biology). View Reference
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Landay A, Gartland GL, Clement LT. Characterization of a phenotypically distinct subpopulation of Leu-2+ cells that suppresses T cell proliferative responses.. J Immunol. 1983; 131(6):2757-61. (Immunogen). View Reference
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Patarroyo M, Makgoba MW. Leucocyte adhesion to cells. Molecular basis, physiological relevance, and abnormalities.. Scand J Immunol. 1989; 30(2):129-64. (Biology). View Reference
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Repo H, Jansson SE, Leirisalo-Repo M. Anticoagulant selection influences flow cytometric determination of CD11b upregulation in vivo and ex vivo.. J Immunol Methods. 1995; 185(1):65-79. (Clone-specific). View Reference
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Ross GD, Cain JA, Lachmann PJ. Membrane complement receptor type three (CR3) has lectin-like properties analogous to bovine conglutinin as functions as a receptor for zymosan and rabbit erythrocytes as well as a receptor for iC3b.. J Immunol. 1985; 134(5):3307-15. (Biology). View Reference
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Shalekoff S, Page-Shipp L, Tiemessen CT. Effects of anticoagulants and temperature on expression of activation markers CD11b and HLA-DR on human leukocytes.. Clin Diagn Lab Immunol. 1998; 5(5):695-702. (Clone-specific). 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.
