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PE Rat Anti-Mouse Dendritic Cells
PE Rat Anti-Mouse Dendritic Cells

Multicolor analysis of splenic dendritic cells. C57BL/6 splenocytes were stained with either PE-conjugated mAb 33D1 (open histograms) or PE-conjugated rat IgG2b, κ isotype control mAb A95-1 (Cat. No. 553989, filled histograms) in the presence of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142). Dendritic cells were identified by staining with APC-conjugated anti-mouse CD11c mAb HL3 (Cat. No. 550261) and FITC-conjugated anti-mouse CD11b mAb M1/70 (Cat. No. 557396/553310). Non-viable leukocytes were excluded by staining with propidium iodide. Left panel displays the expression of CD11c and CD11b among the viable splenocytes, and the gates used for further analysis are shown. The CD11c+CD11b[intermediate] dendritic cell population expresses the 33D1 Dendritic Cell antigen (middle panel), whereas the CD11c-CD11b+ non-dendritic population is 33D1-negative (right panel). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Multicolor analysis of splenic dendritic cells. C57BL/6 splenocytes were stained with either PE-conjugated mAb 33D1 (open histograms) or PE-conjugated rat IgG2b, κ isotype control mAb A95-1 (Cat. No. 553989, filled histograms) in the presence of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142). Dendritic cells were identified by staining with APC-conjugated anti-mouse CD11c mAb HL3 (Cat. No. 550261) and FITC-conjugated anti-mouse CD11b mAb M1/70 (Cat. No. 557396/553310). Non-viable leukocytes were excluded by staining with propidium iodide. Left panel displays the expression of CD11c and CD11b among the viable splenocytes, and the gates used for further analysis are shown. The CD11c+CD11b[intermediate] dendritic cell population expresses the 33D1 Dendritic Cell antigen (middle panel), whereas the CD11c-CD11b+ non-dendritic population is 33D1-negative (right panel). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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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_396754
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

For optimal staining of peripheral leukocytes, we recommend the use of Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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. An isotype control should be used at the same concentration as the antibody of interest.
557578 Rev. 4
Antibody Details
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33D1

The 33D1 antibody reacts with an antigen on most dendritic cells (DC) 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.

557578 Rev. 4
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
557578 Rev.4
Citations & References
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Development References (13)

  1. 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. (Biology). View Reference
  2. 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
  3. 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
  4. 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. (Biology). View Reference
  5. 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. (Biology). View Reference
  6. 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. (Biology). View Reference
  7. 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. (Biology). View Reference
  8. 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). View Reference
  9. 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
  10. 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. (Biology). View Reference
  11. 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. (Biology). View Reference
  12. 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. (Biology). View Reference
  13. Woo J, Lu L, Rao AS, et al. Isolation, phenotype, and allostimulatory activity of mouse liver dendritic cells. Transplantation. 1994; 58(4):484-491. (Biology). View Reference
View All (13) View Less
557578 Rev. 4

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