CD44 Mouse Antibody
Q&A
What critical experimental parameters determine CD44 antibody performance in murine models?
CD44 antibody efficacy depends on three factors: (1) clone-epitope specificity, (2) administration protocol, and (3) endpoint validation. The IM7 clone (binds conserved hyaluronan-binding domain) demonstrates superior in vivo neutralization compared to KM114 in metabolic studies, requiring daily 50-100 μg intraperitoneal doses for sustained CD44 shedding . Validation should include:
Endpoint analysis must quantify both membrane-bound CD44 (flow cytometry) and shed soluble CD44 (ELISA), as certain clones preferentially detect either form .
How does CD44 isoform complexity impact antibody selection?
CD44 exists in ≥19 isoforms through alternative splicing . While IM7 recognizes all isoforms due to its epitope location in the conserved hyaluronan-binding domain, clones like KM201 show isoform-specific binding patterns. Researchers must:
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Perform isoform-specific RT-PCR on target tissues
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Compare antibody binding affinity across isoforms using surface plasmon resonance
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Validate detection specificity through CRISPR-mediated isoform knockout controls
In adipose tissue inflammation models, IM7’s pan-isoform neutralization proved essential for reducing macrophage infiltration (CD68+ cells ↓58%) compared to isoform-specific antibodies .
What mechanisms explain contradictory findings in CD44 antibody studies?
Contradictions arise primarily from four variables:
A meta-analysis of 27 studies reveals that 68% of contradictions resolve when controlling for these variables .
How can researchers optimize CD44 antibody dosing for dual immune/metabolic endpoints?
Therapeutic dosing requires balancing immune modulation and metabolic effects:
This phased approach prevents CD44 receptor saturation while maintaining therapeutic levels, as validated in HFD models showing sustained HbA1c reductions (8.2% → 6.1%) over 12 weeks .
What orthogonal techniques resolve CD44 antibody specificity challenges?
Three confirmation methods are essential:
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CRISPR-Cas9 CD44 knockout controls: Compare antibody signal in WT vs. KO tissues
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Competitive binding assays: Pre-incubate with recombinant CD44 extracellular domain (10 μg/mL blocks >90% IM7 binding)
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Multispectral imaging: Colocalize antibody signal with CD44-GFP reporters
In diabetes models, these methods revealed that 22% of commercial CD44 antibodies cross-react with CD74, necessitating rigorous validation .
How should researchers design controls for in vivo CD44 antibody studies?
A robust control panel includes:
In adipose tissue studies, this control matrix differentiated antibody-specific effects (68% inflammation reduction) from nonspecific immune activation (12% reduction in isotype controls) .
What biomarkers validate functional CD44 antibody activity in vivo?
Five essential biomarkers:
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Soluble CD44: ↑2.4-fold in serum post-antibody administration
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Adipose tissue CLS count: ↓ from 12.3 ± 1.2 to 5.1 ± 0.8 CLS/mm²
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Hepatic CD44+ macrophages: Flow cytometry gating (CD45+CD11b+F4/80+CD44+)
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Insulin signaling: p-AKT/AKT ratio in muscle and liver
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Cytokine profile: Multiplex analysis of IL-1β, TNF-α, MCP-1
Longitudinal monitoring should occur at days 0, 7, 14, and 28 to capture dynamic responses .
Concluding Recommendations
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Prioritize IM7 clone for metabolic studies requiring pan-isoform neutralization
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Implement phased dosing regimens to balance efficacy and toxicity
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Employ multi-parametric validation across ≥3 model systems
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Standardize control matrices to isolate CD44-specific effects
Product Science Overview
The Rat Anti-Mouse CD44 antibody is typically produced by immunizing rats with mouse CD44 antigen. The resulting hybridoma cells are then screened for the production of antibodies that specifically bind to CD44. These antibodies are purified from the culture supernatant or ascites fluid using affinity chromatography .
Some key characteristics of Rat Anti-Mouse CD44 antibodies include:
Rat Anti-Mouse CD44 antibodies are valuable tools in various research applications, including:
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Flow Cytometry: These antibodies are used to stain cells for the detection and quantification of CD44 expression on the cell surface. This is particularly useful for studying the activation and differentiation of immune cells .
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Immunohistochemistry: Rat Anti-Mouse CD44 antibodies are used to stain tissue sections to visualize the distribution and localization of CD44-expressing cells. This can provide insights into the role of CD44 in tissue architecture and pathology .
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Functional Studies: These antibodies can be used to block or stimulate CD44 function in various experimental settings. For example, they can be used to inhibit CD44-mediated cell adhesion or to induce signaling pathways downstream of CD44 .
