Multicolor flow cytometric analysis of mouse splenic dendritic cells. C57BL/6 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 APC Rat Anti-Mouse CD11b (Cat. No. 553312/561690) and BD Horizon™ BV421 Hamster Anti-Mouse CD11c (Cat. No. 562782) antibodies, and either PerCP-Cy™5.5 Rat IgG2b, κ Isotype Control (Cat. No. 550764; dashed line histograms) or PerCP-Cy™5.5 Rat Anti-Mouse Dendritic Cells antibody (Cat. No. 565222; solid line histograms). The fluorescence histogram showing Dendritic Cells antigen expression (or Ig Isotype control staining) were derived from either CD11b+ CD11c- (Left Panel) or CD11b [intermediate] CD11c+ (Right Panel) gated events, as indicated, with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of mouse splenic dendritic cells. C57BL/6 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 APC Rat Anti-Mouse CD11b (Cat. No. 553312/561690) and BD Horizon™ BV421 Hamster Anti-Mouse CD11c (Cat. No. 562782) antibodies, and either PerCP-Cy™5.5 Rat IgG2b, κ Isotype Control (Cat. No. 550764; dashed line histograms) or PerCP-Cy™5.5 Rat Anti-Mouse Dendritic Cells antibody (Cat. No. 565222; solid line histograms). The fluorescence histogram showing Dendritic Cells antigen expression (or Ig Isotype control staining) were derived from either CD11b+ CD11c- (Left Panel) or CD11b [intermediate] CD11c+ (Right Panel) gated events, as indicated, with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
製品詳細
BD Pharmingen™
Dcir2; Dendritic Cell Marker; DC Marker; Dendritic Cells Ag; 33D1 Antigen
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2b, κ
Dendritic cells purified from mouse spleen and lymph node
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739119
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
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.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
- PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
関連製品
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ
0.1 mg
カタログ番号
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ
0.5 mg
カタログ番号
553142
Stain Buffer (FBS) RUO
サイズ
500 mL
カタログ番号
554656
Stain Buffer (BSA) RUO
サイズ
500 mL
カタログ番号
554657
Lysing Buffer RUO
サイズ
100 mL
カタログ番号
555899
PerCP-Cy™5.5 Rat IgG2b, κ Isotype Control RUO
サイズ
0.1 mg
カタログ番号
550764
565222 Rev. 2
抗体の詳細
33D1
The 33D1 antibody recognizes Dendritic cell inhibitory receptor 2 (Dcir2) which is also known as, Dendritic Cells antigen, Dendritic Cell (DC) Marker, or 33D1 antigen. This antigen is expressed on most dendritic cells of spleen, lymph node, and Peyer's patch, but not liver, bone marrow, or epidermal dendritic cells; macrophages; other leukocytes; or erythroid cells. Within the spleen, the majority of 33D1+ DC are localized in the marginal zones. Thymic dendritic cells may express a low level of the 33D1 Antigen. It has been reported that bone-marrow DC can be induced to express the 33D1 antigen by culture in the presence of GM-CSF, and the resulting 33D1+ DC are effective in in vitro (induction of MLR) and in vivo (anti-tumoral vaccination) assays for antigen presentation. However, the addition of IL-4 to GM-CSF in bone-marrow cultures resulted in loss of 33D1 expression and enhanced the MLR-stimulatory activity of the DC. It has also been reported that 33D1 expression is upregulated when liver-derived DC are cultured on collagen-coated plates in the presence of GM-CSF. In vivo functional 33D1+ DC are induced in the brains of mice chronically infected with Toxoplasma gondii, probably via the parasite's induction of GM-CSF.
565222 Rev. 2
フォーマットの詳細
PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
565222 Rev.2
引用＆参考文献
Development References (13)
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Crowley M, Inaba K, Witmer-Pack M, Steinman RM. The cell surface of mouse dendritic cells: FACS analyses of dendritic cells from different tissues including thymus. Cell Immunol. 1989; 118(1):108-125. (Clone-specific: Flow cytometry). View Reference
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Dudziak D, Kamphorst AO, Nussenzweig MC, et al. Differential antigen processing by dendritic cell subsets in vivo. Science. 2007; 315(5808):107-111. (Clone-specific: Flow cytometry). View Reference
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Fischer HG, Bonifas U, Reichmann G. Phenotype and functions of brain dendritic cells emerging during chronic infection of mice with Toxoplasma gondii. J Immunol. 2000; 164(9):4826-4834. (Biology). View Reference
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Inaba K, Inaba M, Romani N, et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J Exp Med. 1992; 176(6):1693-1702. (Clone-specific: Flow cytometry). View Reference
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Kelsall BL, Strober W. Distinct populations of dendritic cells are present in the subepithelial dome and T cell regions of the murine Peyer's patch. J Exp Med. 1996; 183(1):237-247. (Clone-specific: Flow cytometry). View Reference
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Lu L, Woo J, Rao AS, et al. Propagation of dendritic cell progenitors from normal mouse liver using granulocyte/macrophage colony-stimulating factor and their maturational development in the presence of type-1 collagen. J Exp Med. 1994; 179(6):1823-1834. (Clone-specific: Flow cytometry). View Reference
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Masurier C, Pioche-Durieu C, Colombo BM, et al. Immunophenotypical and functional heterogeneity of dendritic cells generated from murine bone marrow cultured with different cytokine combinations: implications for anti-tumoral cell therapy. Immunology. 1999; 96(4):569-577. (Clone-specific: Flow cytometry). View Reference
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Nussenzweig MC, Steinman RM, Witmer MD, Gutchinov B. A monoclonal antibody specific for mouse dendritic cells. Proc Natl Acad Sci U S A. 1982; 79(1):161-165. (Immunogen: Cytotoxicity, Depletion, Radioimmunoassay). View Reference
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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: Flow cytometry). View Reference
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Steinman RM, Gutchinov B, Witmer MD, Nussenzweig MC. Dendritic cells are the principal stimulators of the primary mixed leukocyte reaction in mice. J Exp Med. 1983; 157(2):613-627. (Clone-specific: Cell separation, Cytotoxicity, Functional assay, Immunofluorescence). View Reference
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Vremec D, Zorbas M, Scollay R, et al. The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells. J Exp Med. 1992; 176(1):47-58. (Clone-specific: Flow cytometry). View Reference
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Witmer MD, Steinman RM. The anatomy of peripheral lymphoid organs with emphasis on accessory cells: light-microscopic immunocytochemical studies of mouse spleen, lymph node, and Peyer's patch. Am J Anat. 1984; 170(3):465-481. (Clone-specific: Immunohistochemistry). View Reference
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Woo J, Lu L, Rao AS, et al. Isolation, phenotype, and allostimulatory activity of mouse liver dendritic cells. Transplantation. 1994; 58(4):484-491. (Clone-specific: Flow cytometry). View Reference
565222 Rev. 2
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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.
