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Multiparameter flow cytometric analysis using BD OptiBuild™ RB744 Mouse Anti-Human CD11b antibody (Cat. No. 757193) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
BD OptiBuild™ RB744 Mouse Anti-Human CD11b
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
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.
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: Flow cytometry, Fluorescence activated cell sorting). 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: Blocking, Flow cytometry, Fluorescence activated cell sorting, Immunoprecipitation, Radioimmunoassay). 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: Flow cytometry). 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: Flow cytometry). View Reference
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.