Anti-Mouse CD22 – Purified in vivo GOLD™ Functional Grade

Clone Cy34.1 is a monoclonal antibody that targets mouse CD22, and its most common in vivo applications in mice include:

• B cell depletion (often administered in combination with anti-CD19 or anti-rat κ light chain antibodies).
• Modulation of B cell function, particularly in studies of autoimmune disease, B cell activation, and proliferation.
• Flow cytometry analysis of B cell populations, both for depletion confirmation and immunophenotyping.
• Immunoprecipitation for analyzing B cell signalling complexes and protein-protein interactions.
• Emerging use in neuroimmunology, with roles in studying B cell involvement in CNS immune responses.

Key Details:

• Cy34.1 binds CD22, primarily expressed on mature follicular and marginal zone B lymphocytes, B-1 cells, and plasma cells.
• Used to study the role of B cells in autoimmunity, as CD22 is a negative regulator of B cell activation.
• Can augment B cell proliferation in response to certain stimuli (e.g., LPS or anti-IgM).
• Frequently used alongside other depletion antibodies for more profound B cell removal in murine models.
• Suitable for in vivo functional studies requiring low endotoxin levels and high purity.

Experimental Usage:

• Typical dosing for B cell depletion involves injection, with confirmation by flow cytometry.
• Used in research on immunology, transplantation, lymphoma models, and neuroimmune interactions.

In summary, clone Cy34.1 is a versatile antibody for in vivo B cell depletion, B cell activity modulation, and flow cytometry in mice, with expanding relevance in studies exploring B cell roles beyond classical immunology.

In the literature, Cy34.1, which targets mouse CD22, is often used in combination with other antibodies for various applications, particularly for B cell studies. Here are some commonly used antibodies or proteins that are utilized alongside Cy34.1:

1. Anti-CD19: This antibody, such as clone 1D3, is often used in combination with Cy34.1 for in vivo B cell depletion studies.

2. Anti-rat κ Light Chain: Clone MAR 18.5 is another antibody that is commonly used in conjunction with Cy34.1 for similar B cell depletion studies.

3. Anti-mouse IgM: Donkey F(ab′)2 against mouse IgM is used to identify B cells and their activation status, sometimes in studies related to CD22.

4. Anti-mouse B220 (RA3-6B2): This antibody is used to mark B cells and is often used in flow cytometry studies alongside CD22.

These combinations are used to study B cell dynamics, activation, and depletion in various immune system contexts.

In addition to these antibodies, other proteins such as LPS (Lipopolysaccharide) are used to stimulate B cell proliferation, which can be augmented by the presence of Cy34.1.

Key Findings from Scientific Literature Citing Clone Cy34.1

Antibody Properties and Applications

• Clone Cy34.1 is a monoclonal antibody (mouse IgG1, κ isotype) that exhibits high specificity and sensitivity for mouse CD22, a sialic acid-binding immunoglobulin-like lectin (Siglec-2) expressed primarily on mature B lymphocytes, including follicular, marginal zone, B-1, and plasma cells.
• Fluorescent conjugates (e.g., AF488, PE/Cy5, AF647) of Cy34.1 are widely used for flow cytometry and fluorescence microscopy to study CD22 expression and localization in mouse tissues.
• Suitability for in vivo studies: Purified Cy34.1 is available in functional, low endotoxin grades for in vivo B cell depletion and modulation experiments, often in combination with other B cell-targeting antibodies (e.g., anti-CD19, anti-rat κ Light Chain).

Biological Insights

• CD22 function: CD22 is a negative regulator of B cell activation and helps prevent autoimmune disease by modulating B cell receptor signaling. Loss of CD22 (CD22-deficient mice) results in hyperresponsive B cells with heightened calcium fluxes and proliferation, highlighting its inhibitory role in B cell signaling.
• B cell proliferation: The Cy34.1 antibody augments B cell proliferative responses in vitro when stimulated with LPS or anti-IgM, suggesting that it can modulate or block the inhibitory function of CD22, thereby enhancing B cell activation.
• Alloantigen specificity: Cy34.1 binds CD22 on mouse strains expressing the Lyb-8.2 alloantigen (e.g., C57BL, BALB/c, CBA, C3H/He), but not on strains lacking this alloantigen (e.g., AKR, DBA/1, DBA/2, NZB, PL).
• Association with BCR: CD22 associates with the B cell antigen receptor (BCR) and is phosphorylated upon BCR or MHC class II cross-linking, indicating its involvement in early signaling events.

Therapeutic and Translational Research

• In vivo modulation: Cy34.1 has been used to study the role of CD22 in immune regulation, including its impact on autoimmune disease models and the potential for targeted B cell therapies.
• Myelin debris clearance: Intracerebral injection of Cy34.1 in aged mice has been reported to promote clearance of myelin debris, suggesting a potential application in neuroinflammatory or demyelinating disease research.
• Host defense: CD22 is implicated in host defense mechanisms, such as protection during invasive bacterial infections like pneumococcal disease.

Summary Table: Key Features of Cy34.1-Based CD22 Research

Conclusion

The clone Cy34.1 antibody has become a cornerstone tool in immunology research, enabling precise detection, functional modulation, and depletion of CD22-expressing B cells in mice. Its use has provided critical insights into the negative regulatory functions of CD22 in B cell biology and its relevance in autoimmune diseases, B cell malignancies, and host defense. These findings underscore the importance of CD22 as a therapeutic target and highlight the value of Cy34.1 as a research reagent in both basic and translational immunology.

Dosing regimens for clone Cy34.1 (anti-mouse CD22 monoclonal antibody) vary significantly across different mouse models, as they are adjusted based on experimental objectives, the disease model, administration route, combination therapies, and the specific outcome being measured.

Context and Supporting Details:

• Variation by Experimental Objective and Model: Reports show that dosing regimens depend on whether the aim is B cell depletion, suppression of neuroinflammation, tumor regression, or other preclinical endpoints.
• Doses and Administration Routes:
  • In a neuromyelitis optica spectrum disorder (NMOSD) mouse model, a single intrastriatal injection of 100 μg per mouse was used.
  • In a Raji xenograft mouse model (cancer model using a B cell tumor line), weekly intraperitoneal doses of 10 mg/kg led to 60% tumor regression.
  • A range from 1 to 20 mg/kg per day is mentioned as a general guideline for immunosuppressive monoclonal antibodies in mouse studies, although not all sources specify if this applies directly to Cy34.1.
• Combination Therapies: Cy34.1 is often used in conjunction with other antibodies, such as anti-CD19 (clone 1D3), to enhance B cell depletion efficacy.
• Targeted Mouse Strains: The antibody is reactive with several common laboratory strains (e.g., BALB/c, A), but optimal dosing may still require empirical determination for each model.

Summary Table of Reported Cy34.1 Dosing Regimens in Mouse Models

• Few Standardized Protocols: There is no universally accepted dosing protocol for Cy34.1—regimens are individualized for each application and preclinical study.

• Critical Considerations:

  • Dose optimization may require pilot studies to balance efficacy and off-target or adverse effects for each combination of mouse model and therapeutic goal.
  • Pay careful attention to recommendations in primary literature or product datasheets, as dosing may change with route (e.g., intravenous vs. intraperitoneal vs. intra-brain) and specific experimental context.

In summary, dosing regimens for Cy34.1 are model-dependent, with published doses ranging from 100 μg/mouse (single, CNS delivery) up to at least 10 mg/kg (weekly, systemic), and possible daily regimens for some immunosuppressive scenarios, but empirical optimization is generally required for each specific experimental setup.