Staining of BALB/c splenocytes with 0.25 µg of Rat IgG2b κ Isotype Control Purified (cat. 14-4031) (open histogram) or 0.25 µg of Anti-Mouse Dendritic Cell Marker (33D1) Purified (filled histogram) followed by Anti-Rat IgG Biotin (cat. 13-4813) and Streptavidin PE (cat. 12-4317). Total viable cells were used for analysis.
Contents: Anti-Mouse Dendritic Cell Marker (33D1) Purified Catalog Number: 14-5884 Concentration: 0.5 mg/ml Formulation: Phosphate buffer pH 7.2, 150 mM NaCl, 0.09% NaN3 Storage Conditions: Store at 2-8°C. Clone: 33D1 Host/Isotype: Rat IgG2b, κ
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14-5884-81
50 ug
14-5884-82
100 ug
14-5884-85
500 ug
Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The 33D1 monoclonal antibody reacts with a mouse DC-specific surface marker. The nature and biological activity of the 33D1 antigen is as of yet unknown. 33D1 has been reported on a variety of dendritic cell subpopulations from mouse thymus, spleen, lymph node, and Peyer’s patch. Bone marrow dendritic cells require GM-CSF to express the 33D1 antigen and this expression is down regulated in the presence of IL-4. 33D1 antigen has been detected in vivo in brain dendritic cells post infection via Toxoplasma gondii.
Applications Reported
For research use only, not for diagnostic or therapeutic use. This 33D1 antibody has been reported for use in flow cytometric analysis, immunohistology staining of frozen tissue sections, and immunohistochemical staining. It has also been reported in complement-mediated cytotoxicity. (Please use Functional Grade purified 33D1 in functional assays.)
Applications Tested
The 33D1 antibody has been tested by flow cytometric analysis of mouse splenocyte cell suspensions. This can be used at less than or equal to 1 μg per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Special Notes
For detection of purified 33D1 it is recommended not to use goat anti-rat due to it's low affinity for 33D1. Rather, we recommend using biotin mouse anti-rat (cat. 13-4813).
References
Nussenzweig, M.C., R.M. Steinman, M.D. Witmer, and B. Gutchinov. 1982. A monoclonal antibody specific for mouse dendritic cells. Proc. Natl. Acad. Sci. USA 79: 161 - 165.
Steinman, R.M., B. Gutchinov, M.D. Witmer, and M.C. Nussenzweig. 1983. Dendritic cells are the principal stimulators of the primary mixed leukocyte reaction in mice. J. Exp. Med. 157: 613 - 627.
Inaba, K., M. Inaba, N. Romani, H. Aya, M. Deguchi, S. Ikehara, S. Muramatsu, and R.M. Steinman. 1992. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J. Exp. Med. 176: 1693 - 1702.
Masurier, C., C. Pioche-Durieu, B.M. Colombo, R. Lacave, F.M. Lemoine, D. Klatzmann, and M. Guigon. 1999. Immunophenotypical and functional heterogeneity of dendritic cells generated from murine bone marrow cultured with different cytokine combinations: implications for anti-tumoral cell therapy. Immunology 96: 569 - 577.
Fischer, H.-G., U. Bonifas, and G. Reichmann. 2000. Phenotype and functions of brain dendritic cells emerging during chronic infection of mice with Toxoplasma gondii. J. Immunol. 164: 4826 - 4834.
Dudziak D, Kamphorst AO, et al. 2007. Differential antigen processing by dendritic cell subsets in vivo. Science 315(5808):107-11. (33D1, IHC frozen, PubMed)
