Anti-Mouse Dendritic Cells – Purified in vivo PLATINUM™ Functional Grade
Anti-Mouse Dendritic Cells – Purified in vivo PLATINUM™ Functional Grade
Product No.: D212
Clone 33D1 Target Dendritic Cells Formats AvailableView All Product Type Monoclonal Antibody Alternate Names DC Marker, 33D1, DCIR2 (dendritic cell inhibitory receptor 2) Isotype Rat IgG2b κ Applications in vivo , WB |
Antibody DetailsProduct DetailsReactive Species Mouse Host Species Rat Recommended Isotype Controls Recommended Isotype Controls Recommended Dilution Buffer Immunogen Dendritic cells Product Concentration ≥ 5.0 mg/ml Endotoxin Level <0.5 EU/mg as determined by the LAL method Purity ≥98% monomer by analytical SEC ⋅ >95% by SDS Page Formulation This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration. Product Preparation Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates. Pathogen Testing To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM™ antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile. Storage and Handling Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles. Country of Origin USA Shipping Next Day 2-8°C RRIDAB_2829893 Applications and Recommended Usage? Quality Tested by Leinco FC The suggested concentration for clone 33D1 antibody for staining cells in flow cytometry is ≤ .25 μg per 106 cells in a volume of 100 μl or 100μl of whole blood. Titration of the reagent is recommended for optimal performance for each application. Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change. DescriptionDescriptionSpecificity Clone 33D1 recognizes mouse DCIR2. Background Dendritic cells are antigen presenting cells that have two functions. They scan the body collecting and processing antigen material that they present on the cell surface to T cells, and they maintain T cell tolerance to “self”. The morphology of dendritic cells is characterized by an extremely large surface-to-volume ratio. Murine splenic dendritic cells can occur in two types: myeloid (cDC) and lymphoid (pDC). Lymphoid dendritic cells produce high amounts of IFN-α and are also called Plasmacytoid dendritic cell because they have an appearance similar to plasma cells. Myeloid, or conventional dendritic cells, secrete IL-12, IL-6, TNF, and chemokines and can be further categorized into three subtypes (CD4−CD8+, CD4+CD8− and CD4−CD8−). These differ from other migratory dendritic cells such as Langerhans cells and interstitial dendritic cells that migrate from peripheral tissues to the lymph nodes. The exact nature and biological activity of the dendritic cell surface marker DCIR2 is currently unknown. DCs are known to play a role in several diseases including myeloid cancer, pDC leukemia, HIV, lupus erythematosus, Crohn's disease and ulcerative colitis. However, it is thought that DCs may be able to control cancer progression because increased densities of DC populations have been linked with better clinical outcome. Lung cancers have been found to include four different subsets of dendritic cells; some of which can activate immune cells that can suppress tumor growth. Dendritic cells have also been shown to play a role in the success of cancer immunotherapies in experimental models. Specifically, the immune checkpoint blocker anti-PD-1 has been shown to indirectly activate DCs through IFN-γ released from drug-activated T cells. Agonizing the non-canonical NF-κB pathway also activates DCs and further enhances anti-PD-1 therapy in an IL-12-dependent manner. Antigen Distribution Murine DCIR2 is found on dendritic cells of the thymus, spleen, lymph nodes, and Peyer’s patches. DCs in the bone marrow may express DCIR2 in the presence of GM-CSF. However, this expression is notably downregulated when IL-4 is present. Furthermore, DCIR2 has been found In vivo on brain dendritic cells post infection with T. gondii. Function GM-CSF is reported to increase expression of 33D1 antigen on dendritic cells from bone marrow cells and IL-4 reported to down regulate the 33D1 antigen. PubMed UniProt.org Research Area Immunology Leinco Antibody AdvisorPowered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments. Clone 33D1 is most commonly used in vivo in mice to selectively deplete or track DCIR2+ conventional type 2 dendritic cells (cDC2), which are involved in immune regulation and disease models. This antibody specifically recognizes the DCIR2 (also called Clec4a4) surface marker, which is mainly expressed on cDC2 in mouse spleen, thymus, lymph nodes, Peyer's patches, and, under certain conditions, in the brain. Typical in vivo applications of clone 33D1 include:
Summary Table: Common In Vivo Uses of 33D1
Key points
Alternative applications include ex vivo sorting after in vivo antibody injection and synergy with various disease or inflammation models. Commonly, the 33D1 antibody is used in combination with several other antibodies to refine the identification and isolation of dendritic cell (DC) subsets in mice by flow cytometry or other immunological techniques. The most frequently co-used antibodies and proteins include:
These markers collectively allow precise definition of cDC2 (33D1^+) cells within complex cell mixtures, such as splenocytes or lymph node suspensions. Additional context and usage:
These marker combinations are standard in immunological studies and are recommended in many protocols for accurate dendritic cell subset identification in mouse tissues. Clone 33D1 represents a landmark monoclonal antibody in dendritic cell research, with extensive scientific literature documenting its specificity, functionality, and applications in immunological studies. This rat-derived hybridoma antibody has become an essential tool for identifying and manipulating dendritic cell populations in mice. Specificity and Antigen RecognitionClone 33D1 recognizes a dendritic cell-specific surface antigen now identified as DCIR2 (Dendritic Cell Inhibitory Receptor 2), also known as Clec4a4 (C-type lectin domain family 4, member a4). The original characterization demonstrated that this antibody specifically killed 80-90% of dendritic cells from spleen and lymph node while showing no reactivity with other leukocytes, including Ia+ and Ia- macrophages, lymphocytes, granulocytes, platelets, or erythroid cells. Quantitative binding studies revealed that dendritic cells express approximately 14,000 binding sites per cell for this antibody. The expression pattern is primarily observed in conventional type 2 dendritic cells (cDC2) within mouse thymus, spleen, lymph nodes, and Peyer's patches. An important technical consideration is that binding of clone 33D1 is calcium-dependent, requiring EDTA-free buffers containing Ca2+ ions for optimal staining. Functional ApplicationsThe antibody has proven invaluable for functional studies of dendritic cells through complement-mediated cytotoxicity. When used with rabbit complement, clone 33D1 selectively depletes dendritic cells from heterogeneous cell populations, enabling researchers to assess the functional consequences of dendritic cell removal. Studies using this approach demonstrated that removal of dendritic cells from unfractionated spleen suspensions reduced stimulatory capacity by 75-90% in mixed leukocyte reactions, comparable to results obtained with specific anti-Ia antibody and complement. The antibody effectively ablates both proliferative and cytotoxic T cell responses when dendritic cells are eliminated, confirming that dendritic cells serve as the principal stimulators of primary immune responses. Importantly, the 33D1 antibody itself does not inhibit stimulation by enriched dendritic cell populations; cytotoxic effects require the presence of complement. Biological Significance of DCIR2Recent research has expanded understanding of the DCIR2 antigen recognized by clone 33D1. This protein is a 236 amino acid molecule with a predicted molecular weight of 27.3 kDa that undergoes post-translational modifications including disulfide bond formation, which may result in dimer appearance in immunoblot assays. DCIR2 is involved in multiple immunological processes including antigen recognition, suppression of autoimmunity through down-regulation of T cell priming, modulation of T cell responses, and progression of diet-induced obesity and inflammation. Studies have shown that DCIR2-rich dendritic cells can ameliorate diseases with strong immune inflammatory components, including experimental autoimmune encephalomyelitis (EAE), experimental melanoma, and diabetes. The antibody's ability to target antigens to cDC2s was first demonstrated in 2007, when immunization with αDCIR2-OVA conjugates successfully induced immune responses. Research UtilityClone 33D1 has been extensively used for both in vitro and in vivo depletion of dendritic cells in experimental studies involving mouse models. Its stability as a marker is noteworthy—dendritic cells continue to express the 33D1 antigen after 4 days in culture, while macrophages and lymphocytes do not acquire it. This property, combined with its high specificity, makes clone 33D1 an essential tool for monitoring dendritic cell content in complex lymphoid mixtures and investigating dendritic cell biology in various disease contexts. Dosing regimens for clone 33D1 (anti-mouse DCIR2) can vary significantly depending on the mouse model, experimental objectives, and immune context, but published protocols most frequently use intraperitoneal administration of 0.5 mg per mouse for 3 consecutive days, followed by weekly boosters up to 6 weeks in BALB/c models.
Summary Table: Common 33D1 Dosing Regimens
Additional considerations:
In summary, the most commonly cited regimen for clone 33D1 is 0.5 mg i.p. for 3 days plus weekly boosters, but efficacy and dose may require adjustment in models involving strong immune stimulation, different mouse strains, or specific research goals. References & CitationsSteinman, R. M. et al. (1982) Pro. Natl. Acad. Sci. USA 79:161
Steinman, R. M. et al. (1983) J. Exp. Med. 157:613
Nussenzweig et al. 1982. Proc Natl Acad Sci U S A. 79(1):161-5. PMID: 6948298.
Technical ProtocolsCertificate of Analysis |
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