Anti-Mouse Dendritic Cells – Purified in vivo GOLD™ Functional Grade

Clone 33D1, a monoclonal antibody targeting the dendritic cell marker DCIR2 (also known as C-type lectin domain family 4, member a4 or Clec4a4), is commonly used in vivo in mouse models for several applications:

1. Dendritic Cell Depletion and Tracking: The primary use of clone 33D1 is to selectively deplete or track mouse DCIR2+ dendritic cells, specifically conventional type 2 dendritic cells (cDC2), in vivo. This allows researchers to investigate the role of these cells in immune responses and disease mechanisms .

2. Immunological Studies: By targeting DCIR2, researchers can study the functions of dendritic cells in immune tolerance, antigen presentation, and the modulation of T cell responses. This is valuable for understanding how dendritic cells contribute to autoimmune diseases and inflammation .

3. Disease Modeling: Clone 33D1 is used in models of diseases such as experimental autoimmune encephalomyelitis (EAE), melanoma, and diabetes, where dendritic cells play a critical role in disease progression and immune response modulation .

4. Research on Obesity and Inflammation: DCIR2-expressing dendritic cells have been implicated in diet-induced obesity and inflammation, making clone 33D1 useful for studying these conditions in mouse models .

Overall, clone 33D1 provides a powerful tool for dissecting the complex roles of dendritic cells in the immune system, offering insights into their functions and potential therapeutic targets.

In the literature, several antibodies and proteins are commonly used alongside 33D1, which targets dendritic cells, particularly conventional type 2 dendritic cells (cDC2). Here are some of these commonly used antibodies and proteins:

• CD11c: A pan-dendritic cell marker, often used to identify dendritic cells broadly.
• CD8α (Ly6G): A marker for cDC1, which helps differentiate between cDC1 and cDC2.
• F4/80: Used to exclude macrophages from the population, as it is a marker for macrophages.
• MHC class II: Utilized to confirm the antigen-presenting capability of dendritic cells.
• B220: Excludes plasmacytoid dendritic cells (pDCs).
• CD45: Helps in gating leukocytes during flow cytometry.

These antibodies help in defining the specific type of dendritic cells and excluding other cell types, which is crucial for studying dendritic cell functions and their role in immune responses. The combination of these markers provides a comprehensive view of the dendritic cell population in various tissues and experimental conditions.

The key findings from scientific literature using clone 33D1 citations are:

• Clone 33D1 specifically recognizes a mouse dendritic cell antigen now known as DCIR2 (Dendritic Cell Inhibitory Receptor 2, also called Clec4a4), a 236 amino acid protein highly expressed on conventional type 2 dendritic cells (cDC2) in the mouse.

• 33D1 antibody provides a reliable and specific marker for murine dendritic cells in tissues such as spleen, lymph node, thymus, and Peyer’s patch. It binds to approximately 14,000 sites per dendritic cell and does not cross-react with macrophages, lymphocytes, granulocytes, platelets, or erythroid cells.

• Functional studies using 33D1 have demonstrated its utility in selectively depleting dendritic cells in vivo and in vitro, allowing analysis of dendritic cell function in immune responses. Complement-mediated killing with 33D1 specifically removes DCs, resulting in ablated T cell responses in mixed leukocyte reactions.

• DCIR2/33D1+ dendritic cells are involved in antigen presentation, regulation of T cell priming, and modulation of immune responses. Removal or depletion of these cells has been shown to suppress both the proliferation and cytotoxic capacity of T cells.

• Recent work identifies DCIR2+ DCs (33D1+ cells) as regulators of autoimmunity, inflammation, and metabolic pathways, and their presence can ameliorate inflammation in disease models such as experimental autoimmune encephalomyelitis, melanoma, and diabetes in mice.

• The molecular and biological functions of the DCIR2/33D1 antigen are still under investigation, but its role in immune cell communication, especially suppression of autoimmunity and T cell priming, is increasingly recognized.

• Clone 33D1 has become a widely adopted reagent for both basic and disease model research concerning dendritic cell function, subset identification, and cell depletion in murine systems.

These findings establish clone 33D1 as an essential tool for immunological studies focused on dendritic cell biology, immune regulation, and related disease mechanisms.

Dosing regimens for clone 33D1 (anti-mouse DCIR2 antibody) vary depending on the mouse model and experimental goal, but several established protocols provide guidance:

• In BALB/c mice, a common regimen for depleting 33D1+ dendritic cells involves intraperitoneal (i.p.) injection of 0.5 mg (500 µg) purified anti-33D1 mAb daily for 3 consecutive days, followed by weekly booster injections for up to 5 weeks to maintain depletion over extended periods. This approach enables near-complete and sustained removal of 33D1+ DCs from the spleen.

• When using single-dose strategies in other experimental settings, some studies have used 100 µg per mouse i.p., administered 48 hours before analysis or additional interventions. This is typically for acute effects or shorter depletion windows.

• According to general antibody dosing overviews for mouse models (while not 33D1-specific), doses in the range of 100–500 µg per mouse i.p. are common for in vivo monoclonal antibody experiments, with dosing frequency adapted to the antibody's half-life and experimental timeline.

Model-specific considerations:

• In allergic airway (AR) models, weekly boosting after the initial triple dose is necessary for continuous depletion, especially because adjuvants like alum/OVA can partially sustain 33D1+ DC numbers even under antibody pressure.
• Different mouse strains or disease models may require slight adjustments to regimen, but the above dosing schemes are widely referenced in immunology literature.

Summary Table: 33D1 Dosing Regimens in Mouse Models

Key points:

• Sustained depletion requires initial multi-day dosing plus weekly maintenance.
• Acute depletion may use a single dose 1–2 days before endpoint assessment.
• Adjuvants (e.g., alum) may reduce depletion efficacy, requiring regimen adjustment.
• Dosing can be adapted for other strains, but 100–500 µg/mouse i.p. is the common range.

If you need dosing protocols for a specific mouse strain or unique disease model, please specify, as some regimens may require further adjustment.