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Purified Mouse Anti-Human CD11c
Purified Mouse Anti-Human CD11c

Analysis of CD11c Expression

     Left Panel - Flow cytometric analysis of CD11c expression on human peripheral blood leucocytes. Human whole blood was (collected with heparin as the preferred anticoagulant rather than EDTA) stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; Left Plot) or Purified Mouse Anti-Human CD11c  antibody (Cat. No. 565805; Right Plot). After washing, the cells were secondarily stained with PE Goat anti-Mouse Ig (Cat. No. 550589). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD11c (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

     Right Panel - Three-color Immunofluorescense analysis of CD11c expression by cells in human tonsil. A human tonsil cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1× PBS, and stained with Purified Mouse Anti-Human CD11c antibody followed by secondary staining with BD Horizon™ BV480 Goat Anti-Mouse Ig (Cat. No. 564877, pseudocolored green). The section was thoroughly washed, then stained with BD Horizon BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427, pseudocolored red) and Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697, pseudocolored blue). The slide was mounted with ProLong® Gold. The images were captured using a standard epifluorescence microscope and merged. Original magnification, 20×.

Analysis of CD11c Expression

     Left Panel - Flow cytometric analysis of CD11c expression on human peripheral blood leucocytes. Human whole blood was (collected with heparin as the preferred anticoagulant rather than EDTA) stained with either Purified Mouse IgG1, κ Isotype Control (Cat. No. 554121; Left Plot) or Purified Mouse Anti-Human CD11c  antibody (Cat. No. 565805; Right Plot). After washing, the cells were secondarily stained with PE Goat anti-Mouse Ig (Cat. No. 550589). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD11c (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.

     Right Panel - Three-color Immunofluorescense analysis of CD11c expression by cells in human tonsil. A human tonsil cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1× PBS, and stained with Purified Mouse Anti-Human CD11c antibody followed by secondary staining with BD Horizon™ BV480 Goat Anti-Mouse Ig (Cat. No. 564877, pseudocolored green). The section was thoroughly washed, then stained with BD Horizon BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427, pseudocolored red) and Alexa Fluor® 488 Mouse Anti-Human CD19 antibody (Cat. No. 557697, pseudocolored blue). The slide was mounted with ProLong® Gold. The images were captured using a standard epifluorescence microscope and merged. Original magnification, 20×.

Product Details
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BD Pharmingen™
ITGAX; AlphaX Integrin; Axb2; Integrin alpha-X; CR4; SLEB6; p150,95 alpha
Human (QC Testing), Rhesus (Tested in Development)
Mouse IgG1, κ
Human monocytes and synovial cells
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.5 mg/ml
III 278; IV M66
3687
AB_2739363
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Note: The binding of the 3.9 antibody to CD11c is divalent cation dependent. Therefore, heparin is recommended for use as the blood anticoagulant rather than the EDTA chelating agent that might adversely affect 3.9 antibody binding and cellular staining

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. 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 affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565805 Rev. 3
Antibody Details
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3.9

The 3.9 monoclonal antibody specifically binds to CD11c, which is also known as Integrin alpha X (αX Integrin/ITGAX), or p150,95 Integrin alpha chain. CD11c is a ~150 kDa type I transmembrane glycoprotein. It is expressed on monocytes, macrophages, granulocytes, NK cells, dendritic cells, and subsets of B and T cells. It associates with CD18 (Integrin beta 2/β2 Integrin) to form the CD11c/CD18 complex, which is also known as p150,95 Integrin, or the Type 4 Complement Receptor (CR4). CD11c/CD18 binds fibrinogen and reportedly serves as a receptor for iC3b and ICAM-1/CD54. CD11c/CD18 functions as an adhesion molecule that mediates cellular binding to ligands expressed on stimulated epithelium and endothelium. The 3.9 monoclonal antibody crossreacts with CD11c expressed by Rhesus macaque leucocytes.

565805 Rev. 3
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
565805 Rev.3
Citations & References
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Development References (7)

  1. Autissier P, Soulas C, Burdo TH, Williams KC. Immunophenotyping of lymphocyte, monocyte and dendritic cell subsets in normal rhesus macaques by 12-color flow cytometry: clarification on DC heterogeneity.. J Immunol Methods. 2010; 360(1-2):119-28. (Clone-specific: Flow cytometry). View Reference
  2. Hogg N, Horton MA. Myeloid antigens: New and previously defined clusters. In: McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:576-602.
  3. Hogg N, Takacs L, Palmer DG, Selvendran Y, Allen C.. The p150,95 molecule is a marker of human mononuclear phagocytes: comparison with expression of class II molecules.. Eur J Immunol. 1986; 16(3):240-248. (Immunogen: Flow cytometry, Immunohistochemistry, Immunoprecipitation). View Reference
  4. Myones BL, Dalzell JG, Hogg N, Ross GD. Neutrophil and monocyte cell surface p150,95 has iC3b-receptor (CR4) activity resembling CR3.. J Clin Invest. 1988; 82(2):640-51. (Clone-specific: Blocking, Functional assay, Immunohistochemistry, Inhibition, Radioimmunoassay). View Reference
  5. Schmidt RE. Non-lineage/natural killer section report: new and previously defined clusters. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:517-542.
  6. Stain C, Jager U, Majdic O, et al. The phenotyping of human basophils with the Myeloid Workshop Panel. In: McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:720-722.
  7. Van der Schoot CE, Daams M, Von dem Borne AEG, et al. Biochemical analysis of the myeloid panel. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:868-876.
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565805 Rev. 3

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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.