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BV421 Hamster Anti-Mouse CD11c
BV421 Hamster Anti-Mouse CD11c

Flow cytometric analysis of CD11c expression on mouse dendritic cells. BALB/c mouse splenocytes were cultured with recombinant mouse GM-CSF (Cat. No.554586; 5 ng/ml) overnight. The cells were then harvested and stained either with a BD Horizon™ BV421 Hamster IgG1, λ1 Isotype Control (Cat. No. 562919; dashed line histogram) or with the BD Horizon™ BV421 Hamster anti-Mouse CD11c antibody (Cat. No. 562782; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable dendritic cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Immunohistofluorescent analysis of CD11c expression by cells within C57BL/6 mouse spleen. A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Horizon™ BV421 Hamster Anti-Mouse CD11c antibody (Cat. No. 562782, pseudo-colored red), BD Horizon™ BV480 Rat-anti-Mouse CD3 Molecular Complex antibody (Cat. No. 565642, pseudo-colored green), and Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557683, pseudo-colored blue). Images were captured on a standard epifluorescence. Original magnification, 20x.

Flow cytometric analysis of CD11c expression on mouse dendritic cells. BALB/c mouse splenocytes were cultured with recombinant mouse GM-CSF (Cat. No.554586; 5 ng/ml) overnight. The cells were then harvested and stained either with a BD Horizon™ BV421 Hamster IgG1, λ1 Isotype Control (Cat. No. 562919; dashed line histogram) or with the BD Horizon™ BV421 Hamster anti-Mouse CD11c antibody (Cat. No. 562782; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable dendritic cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

Immunohistofluorescent analysis of CD11c expression by cells within C57BL/6 mouse spleen. A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Horizon™ BV421 Hamster Anti-Mouse CD11c antibody (Cat. No. 562782, pseudo-colored red), BD Horizon™ BV480 Rat-anti-Mouse CD3 Molecular Complex antibody (Cat. No. 565642, pseudo-colored green), and Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557683, pseudo-colored blue). Images were captured on a standard epifluorescence. Original magnification, 20x.

Product Details
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BD Horizon™
Cd11c; Itgax; Integrin alpha-X; Integrin αX; Cr4; Complement receptor 4
Mouse (QC Testing)
Armenian Hamster IgG1, λ2
C57BL/6 Mouse Intestinal Intraepithelial Lymphocytes
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.2 mg/ml
16411
AB_2737789
Aqueous buffered solution containing BSA and ≤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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

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).

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  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. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  10. 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.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562782 Rev. 2
Antibody Details
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HL3

The HL3 monoclonal antibody specifically binds to the integrin αx chain of gp150, 95 (CD11c/CD18). CD11c is expressed on dendritic cells, CD4- CD8+ intestinal intraepithelial lymphocytes (IEL) and some NK cells. It is upregulated on IEL and lymph-node T cells following in vivo activation. Cells of the monocyte/macrophage lineage have been reported to express low levels of CD11c. CD11c plays a role in binding of iC3b.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562782 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562782 Rev.2
Citations & References
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Development References (13)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. Burt BM, Plitas G, Stableford JA, et al. CD11c identifies a subset of murine liver natural killer cells that responds to adenoviral hepatitis. J Leukoc Biol. 2008; 84(4):1039-1046. (Clone-specific: Flow cytometry). View Reference
  3. Burt BM, Plitas G, Stableford JA, et al. CD11c identifies a subset of murine liver natural killer cells that responds to adenoviral hepatitis. J Leukoc Biol. 2008; 84(4):1039-1046. (Clone-specific: Flow cytometry, Immunoprecipitation). View Reference
  4. Fagarasan S, Muramatsu M, Suzuki K, Nagaoka H, Hiai H, Honjo T. Critical roles of activation-induced cytidine deaminase in the homeostasis of gut flora. Science. 2002; 298(5597):1424-1427. (Clone-specific: Immunofluorescence). View Reference
  5. Fagarasan S, Muramatsu M, Suzuki K, Nagaoka H, Hiai H, Honjo T. Critical roles of activation-induced cytidine deaminase in the homeostasis of gut flora. Science. 2002; 298(5597):1424-1427. (Clone-specific: Immunofluorescence). View Reference
  6. Gao JX, Liu X, Wen J, et al. Differentiation of monocytic cell clones into CD8 alpha+ dendritic cells (DC) suggests that monocytes can be direct precursors for both CD8 alpha+ and CD8 alpha- DC in the mouse. J Immunol. 2003; 170(12):5927-5935. (Biology). View Reference
  7. Huleatt JW, Lefrançois L. Antigen-driven induction of CD11c on intestinal intraepithelial lymphocytes and CD8+ T cells in vivo.. J Immunol. 1995; 154(11):5684-93. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
  8. Huleatt JW, Lefrançois L. Antigen-driven induction of CD11c on intestinal intraepithelial lymphocytes and CD8+ T cells in vivo.. J Immunol. 1995; 154(11):5684-93. (Immunogen: Immunoprecipitation). View Reference
  9. Larson RS, Springer TA. Structure and function of leukocyte integrins. Immunol Rev. 1990; 114:181-217. (Biology). View Reference
  10. Maraskovsky E, Brasel K, Teepe M, et al. Dramatic increase in the numbers of functionally mature dendritic cells in Flt3 ligand-treated mice: multiple dendritic cell subpopulations identified. J Exp Med. 1996; 184(5):1953-1962. (Biology). View Reference
  11. 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. (Biology). View Reference
  12. Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. (Biology). View Reference
  13. Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. (Clone-specific: Immunohistochemistry). View Reference
View All (13) View Less
562782 Rev. 2

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