Multiparameter flow cytometric analysis of CD11c expression on human peripheral blood leucocyte populations. Human whole blood (collected with heparin as the preferred anticoagulant rather than EDTA) was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CD11c antibody (Cat. No. 565911/565912; Right Plot) at 1 µg/test. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter pseudocolor dot 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 FACSCelesta™ Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD11c expression on human peripheral blood leucocyte populations. Human whole blood (collected with heparin as the preferred anticoagulant rather than EDTA) was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CD11c antibody (Cat. No. 565911/565912; Right Plot) at 1 µg/test. Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter pseudocolor dot 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 FACSCelesta™ Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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)
0.2 mg/ml
III 278; IV M66
3687
AB_2869733
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
50 µg
Cat No.
565571
Alexa Fluor® 647 Mouse Anti-Human CD11c RUO
Size
25 µg
Cat No.
565911
565912 Rev. 1
Antibody Details
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.
565912 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
565912 Rev.1
Citations & References
View product citations for antibody "565912" on CiteAb
Development References (8)
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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
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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.
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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
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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
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Sadhu C, Hendrickson L, Dick KO, Potter TG, Staunton DE. Novel tools for functional analysis of CD11c: activation-specific, activation-independent, and activating antibodies.. J Immunoassay Immunochem. 2008; 29(1):42-57. (Clone-specific: Flow cytometry). View Reference
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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.
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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.
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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.
565912 Rev. 1
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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.