Anti-Mouse IL-21R [Clone 4A9] — Purified in vivo GOLD™ Functional Grade

Clone 4A9 is most commonly used in vivo in mice as a blocking antibody against mouse Interleukin-21 Receptor (IL-21R, also known as CD360) to inhibit IL-21-mediated signaling pathways.

Key in vivo applications include:

• Blocking IL-21R signaling: Direct administration of clone 4A9 in mice is used to block the biological activity of IL-21R, allowing researchers to investigate the functional role of IL-21 signaling in various immune processes and disease models. This is particularly relevant in immunology studies involving T cells, B cells, and NK cells, where IL-21 signaling is known to regulate proliferation, differentiation, and function.
• Investigating disease mechanisms: Clone 4A9 is applied in studies of autoimmune disease, chronic infection, immunoglobulin production, and cancer models to modulate IL-21 activity and study its impact on disease progression.
• Functional immune modulation: By blocking IL-21R, clone 4A9 enables the assessment of IL-21's effects on immune regulation, such as T cell survival, B cell maturation, and antiviral responses.

Additional validated uses (in support of, but not exclusive to in vivo work):

• Clone 4A9 is suitable for other research assays such as flow cytometry, immunoprecipitation, and ELISA, but its low-endotoxin, high-purity preparations are specifically designed for safe and effective in vivo administration.

Summary Table: Main In Vivo Applications of Clone 4A9 in Mice

Clone 4A9 is prepared with ultra-low endotoxin levels to ensure compatibility with sensitive in vivo mouse studies and minimize off-target responses. The antibody is typically a rat IgG2a isotype, and is administered using standard in vivo antibody techniques for preclinical research.

Commonly, 4A9 refers to the mouse monoclonal antibody targeting Aldolase C, a glial cytosolic protein. In the literature, the most frequently used antibodies or proteins together with 4A9 are:

• MAP2 antibody: Often co-stained with 4A9 in immunohistochemistry to differentiate neurons (MAP2 labels dendrites and neuronal cell bodies) from glial Aldolase C-positive cells.
• Vimentin antibody: Used as a glial marker in combination with 4A9, particularly in brain sections; 4A9+ Purkinje cells do not express vimentin, which allows for clear identification.
• DAPI: A fluorescent nuclear stain, routinely used for counterstaining cell nuclei in sections stained with 4A9 and other antibodies.

Other commonly used antibodies in studies involving Aldolase C and/or neural cell identification may include:

• GFAP: A standard marker of astrocytes, sometimes used to define glial populations in combination with Aldolase C (though not specifically mentioned in the provided results, this is widely established in neurobiology).
• Secondary antibodies (HRP, Alexa Fluor-conjugates): Utilized for detection of 4A9 in Western blot, immunocytochemistry, or immunofluorescence workflows.

Key proteins and markers routinely paired with 4A9 in neural tissue studies:

• Neuronal markers: MAP2, NeuN, Synaptophysin, βIII Tubulin, to contrast with Aldolase C-positive glia.
• Glial markers: Vimentin, GFAP.

Applications and detection methods:

• Immunohistochemistry (IHC): Costaining with MAP2, vimentin, and nuclear markers.
• Western blot: May include lysates probed for other metabolic enzymes or neuronal/glial proteins for comparative quantification.
• Immunofluorescence: Co-detection with fluorescently-labeled secondary antibodies and nuclear counterstains.

In summary, Aldolase C antibody clone 4A9 is frequently used alongside MAP2 and Vimentin antibodies, as well as nuclear stains like DAPI, to distinguish neuronal and glial populations during neuroanatomical analyses in rodent tissue.

Clone 4A9 is most prominently referenced in scientific literature as a monoclonal antibody targeting immune receptors in mouse models, specifically anti-mouse IL-21R and anti-mouse Mincle.

Key findings from citations involving clone 4A9 in the literature include:

• Anti-mouse IL-21R (clone 4A9):

  • Clone 4A9 was generated by immunizing Lewis rats with IL-21R-transfected cells and selectively binds mouse IL-21R, as demonstrated by FACS analysis and immunoprecipitation.
  • It stains the majority of spleen cells from wild type C57BL/6 mice, but not from IL-21R-deficient mice, confirming its specificity for IL-21R.
  • IL-21, the ligand, blocks 4A9’s binding to IL-21R-transfected cells, confirming that 4A9 recognizes the ligand-binding domain of IL-21R.
  • IL-21R is broadly expressed on B cells and, to a lesser extent, on T and NK cells in mice.
  • Research using 4A9 has enabled characterizing the function and regulation of IL-21R in immunity, as cited in peer-reviewed studies (e.g., Schmitz et al., PLoS Pathog. 2013; Simard et al., J Immunol. 2011; Jin et al., J Immunol. 2004; DeKoter et al., J Immunol. 2010).

• Anti-mouse Mincle (clone 4A9):

  • In separate literature, clone 4A9 is also described as a monoclonal antibody against mouse Mincle, a receptor critical for the recognition of mycobacterial glycolipids, notably trehalose-6,6'-dimycolate (TDM).
  • Studies using this clone have shown that Mincle is essential for cytokine and chemokine production in response to these glycolipids, implicating its role in antimycobacterial immunity.

These antibodies have enabled precise characterization of immune cell subsets, receptor function, and ligand/receptor interactions, which are foundational for immunology research in mouse models. No evidence from search results indicates off-target reactivity or major controversies in the scientific use of clone 4A9.

If you require findings related to a specific disease model, antigen, or application, please clarify for a more targeted synthesis. Current literature identifies the clone as central to immunophenotyping and functional studies of mouse IL-21R and Mincle.

Dosing regimens for clone 4A9 (monoclonal antibody targeting either Mincle or IL-21R, depending on context) are not standardized across different mouse models; instead, they are highly dependent on the experimental application (e.g., in vitro assays, flow cytometry, in vivo blockade) rather than the mouse strain or disease context.

Key findings on dosing regimens:

• No universally standardized regimen: Available sources explicitly state that published protocols use concentrations based on assay type (functional blocking, detection, immunoprecipitation, etc.), not on the specific mouse strain or disease model.
• In vivo blockade (IL-21R target): For anti–IL-21R (clone 4A9), in a mouse immunization study, two distinct dosing regimens were reported:
  • 25 μg/day, administered from days 7 to 9 after immunization (a low dose).
  • 500 μg/day (a high dose, referenced from prior literature), for three consecutive days.
• In vitro and ex vivo use (detection/blocking):
  • Flow cytometry: 5–10 μg/mL final antibody concentration.
  • Western blot: 1–5 μg/mL.

Variation across models:

• Studies specifically state that regimens do not depend on the mouse strain (e.g., BALB/c, C57BL/6) or disease model (e.g., EAE, tumor, infection).
• Differences arise exclusively based on:
  • The purpose of the experiment (e.g., receptor blocking, staining).
  • The delivery route (e.g., intraperitoneal injection for in vivo, direct addition to staining buffers for ex vivo).
• No comprehensive data show systematic adjustment of dose by genotype, age, sex, or immunological status of the mice.

Summary Table: Clone 4A9 Dosing Contexts

In summary: The dosing regimen for clone 4A9 is dictated by experimental objective, not mouse model background, and a range of concentrations is applied depending on the assay, with documented in vivo doses for IL-21R blockade ranging from 25 μg/day to 500 μg/day for three days, but no guidance for further model-specific adjustment.