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BB515 Armenian Hamster Anti-Mouse CD11c
BB515 Armenian Hamster Anti-Mouse CD11c
Two-color flow cytometric analysis of CD11c expression on C57BL/6 mouse splenic leucocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse I-A/I-E antibody (Cat. No. 562564) and BD Horizon™ BB515 Armenian Hamster Anti-Mouse CD11c antibody (Cat. No. 565586) at 0.25 µg/test. The two-color flow cytometric contour plot showing the correlated expression of CD11c versus I-A/I-E was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of CD11c expression on C57BL/6 mouse splenic leucocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse I-A/I-E antibody (Cat. No. 562564) and BD Horizon™ BB515 Armenian Hamster Anti-Mouse CD11c antibody (Cat. No. 565586) at 0.25 µg/test. The two-color flow cytometric contour plot showing the correlated expression of CD11c versus I-A/I-E was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. 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
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BD Horizon™
Itgax; Integrin alpha-X; Integrin αX; ITAX; CR4; Complement Receptor 4
Mouse (QC Testing)
Armenian Hamster IgG2
Mouse Dendritic Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869690
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 with BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
  6. 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.
  7. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565586 Rev. 2
Antibody Details
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N418

The N418 monoclonal antibody specifically binds to CD11c, the Integrin alpha X (Integrin αX, Itgax) chain of the heterodimeric gp150, 95 (CD11c/CD18, αXβ2) integrin that forms the complement receptor 4 (CR4). CD11c is a150 kDa type I transmembrane glycoprotein that is expressed on dendritic cells, CD4-CD8+ intestinal intraepithelial lymphocytes (IEL), and some NK cells and T cells. CD11c expression is upregulated on IEL and T cells following activation. Cells of the monocyte/macrophage lineage have been reported to express low levels of CD11c. The CD11c/CD18 integrin can bind to several ligands including iC3b, fibrinogen, and CD54. This integrin reportedly plays important roles in phagocytosis and in mediating cellular interactions during inflammation.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

565586 Rev. 2
Format Details
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
565586 Rev.2
Citations & References
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Development References (3)

  1. Crowley MT, Inaba K, Witmer-Pack MD, Gezelter S, Steinman RM. Use of the fluorescence activated cell sorter to enrich dendritic cells from mouse spleen. J Immunol Methods. 1990; 133(1):55-66. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  2. Metlay JP, Witmer-Pack MD, Agger R, Crowley MT, Lawless D, Steinman RM. The distinct leukocyte integrins of mouse spleen dendritic cells as identified with new hamster monoclonal antibodies. J Exp Med. 1990; 171(5):1753-1771. (Immunogen: Flow cytometry, Immunohistochemistry, Immunoprecipitation). View Reference
  3. Sadhu C, Ting HJ, Lipsky B, et al. CD11c/CD18: novel ligands and a role in delayed-type hypersensitivity. J Leukoc Biol. 2007; 81(6):1395-1403. (Clone-specific: Blocking). View Reference
565586 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.