Anti-Mouse CD54 (Clone YN1/1.7.4) – Purified in vivo GOLD™ Functional Grade

Clone YN1/1.7.4, a rat monoclonal antibody against mouse CD54 (ICAM-1), is extensively used in in vivo research to modulate immune responses and inflammation in mice. This antibody has been specifically engineered for in vivo studies with ultra-low endotoxin levels and high purity to ensure reliable experimental outcomes.

Blocking Adhesion and Immune Cell Recruitment

The primary in vivo application involves blocking CD54-mediated cell adhesion and leukocyte trafficking. CD54 binds to integrins LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18) on leukocytes, facilitating their adhesion to endothelial cells and subsequent transmigration into tissues. The YN1/1.7.4 antibody has been shown to block binding of mouse CD54 to both LFA-1 and Mac-1, thereby inhibiting cell-cell adhesion.

Suppressing Inflammatory Responses

In vivo studies demonstrate that YN1/1.7.4 effectively neutralizes CD54 and suppresses inflammatory mediators. The antibody has been shown to inhibit production of CXCL1, IFNγ, and IL-17, which are key inflammatory cytokines. Additionally, it reduces neutrophil infiltration into inflammatory lesions, making it valuable for studying acute inflammatory responses.

Modulating Antigen Presentation

The antibody can also interfere with antigen presentation processes where CD54 plays a role. By blocking CD54 function, researchers can investigate the molecule's contribution to T cell activation and immune cell interactions during adaptive immune responses.

Technical Specifications for In Vivo Use

The in vivo PLATINUM™ formulations of YN1/1.7.4 meet stringent quality standards essential for animal studies. These include endotoxin levels less than 0.001 ng/µg antibody, greater than 90% purity, and less than 10% aggregation. The antibody is provided in sterile, 0.2 µm filtered formulations in phosphate-buffered saline, optimized for direct in vivo administration.

These applications make clone YN1/1.7.4 a valuable tool for investigating inflammatory diseases, immune cell migration, and CD54-dependent pathological processes in mouse models.

Antibodies or proteins commonly used with YN1/1.7.4 (anti-mouse CD54/ICAM-1) in the literature include a variety of markers to identify and characterize immune cell populations and endothelial biology.

Key antibodies and proteins frequently used alongside YN1/1.7.4:

• CD3: T cell marker
• CD4: Helper T cell marker
• CD8: Cytotoxic T cell marker
• CD19: B cell marker
• CD11b: Myeloid/monocyte marker
• CD11c: Dendritic cell marker

Experimental setups often use these markers in flow cytometry panels or immunohistochemistry to differentiate cell types, assess cell activation, or analyze tissue composition.

Other endothelial and leukocyte adhesion markers, notably:

• CD31 (also known as PECAM-1): Another endothelial marker frequently co-stained with CD54 for vascular/endothelial research. Combining anti-CD54 (YN1/1.7.4) and anti-CD31 antibodies is used to improve visualization and analysis of endothelial boundaries, particularly in imaging-based approaches such as confocal laser endomicroscopy.

Additionally, in functional assays related to leukocyte adhesion and migration, LFA-1 (CD11a/CD18, integrin αLβ2) and Mac-1 (CD11b/CD18, integrin αMβ2)—both ligands of ICAM-1—are often relevant for blocking or mechanistic studies, though these are more often proteins of interest than antibodies used in co-labeling.

In summary, studies routinely pair YN1/1.7.4 with:

• Major leukocyte surface markers (CD3, CD4, CD8, CD19, CD11b, CD11c) for immune phenotyping.
• Other endothelial markers like CD31 for vasculature analysis, particularly in imaging applications.
• Sometimes isotype controls and fluorophore-labeled secondary antibodies, depending on the assay.
• Functional blocking studies may investigate interactions with LFA-1 and Mac-1, though not necessarily as antibody pairs.

These combinations enable accurate identification of immune subsets, study of cell trafficking, and characterization of endothelial and inflammatory states.

Key findings from scientific literature citations of clone YN1/1.7.4 center on its use as an anti-mouse CD54 (ICAM-1) monoclonal antibody for blocking, imaging, and characterizing cell adhesion molecule functions.

Essential context and supporting details:

• Functional Blockade: YN1/1.7.4 efficiently blocks CD54 (ICAM-1) interactions with integrin ligands such as LFA-1 (CD11a/CD18) and Mac-1 (CD11b/CD18), inhibiting cell–cell adhesion in endothelial and immune contexts. This blockade is widely used to dissect the role of ICAM-1 in inflammatory and immunological processes.

• Binding and Imaging Applications: The clone is highly specific for mouse CD54 (ICAM-1) and is routinely utilized in flow cytometry, immunohistochemistry, and fluorescent imaging to detect ICAM-1 expression on various cell types (e.g., endothelial cells, lymphocytes, dendritic cells, monocytes). Its specificity has been confirmed for both functional assays and imaging.

• Epitopic Specificity: YN1/1.7.4 recognizes a unique epitope on mouse CD54 different from other antibodies, ensuring reliable functional and imaging distinctions across experiments.

• Immunological Research Utility: Researchers use YN1/1.7.4 to study:

  • Cell adhesion and transmigration by blocking ICAM-1-mediated interactions with leukocytes.
  • The dynamics of immune cell recruitment and trafficking, notably in inflammation, infection, and cancer models.
  • ICAM-1’s role in endothelial barrier function and the response to inflammatory stimuli.

• Imaging and Quantification: In advanced imaging studies, such as fluorescence-guided fiber-optic micronavigation, YN1/1.7.4 enables real-time visualization and quantification of ICAM-1 expression and function on endothelial cells and tissue segments.

• Reported Experimental Protocols:

  • Flow cytometry: Effective staining concentrations are typically ≤0.5 µg/test, titrated for optimal signal.
  • Blocking studies: Doses vary with application, but examples include 50 μg for in vitro blocking and up to several mg/kg for in vivo blockade.
  • Imaging protocols: Used with conjugated fluorophores for high-resolution visualization of cell-surface ICAM-1.

• Cellular Expression Details: ICAM-1 is expressed at variable levels—upregulated especially during inflammation—on endothelial cells, immune cells (lymphocytes, monocytes, macrophages, neutrophils), and some carcinoma and melanoma cells, making YN1/1.7.4 widely applicable in disease models.

Additional relevant information:

• YN1/1.7.4 is frequently cited across immunology, cell biology, inflammation, and oncology research, with applications in both murine and rat models.
• The antibody’s functional grade preparations are critical for blocking assays, while different conjugates enable versatile imaging options.
• Literature suggests careful titration and selection of antibody format (functional grade vs. imaging conjugate) for precise experimental outcomes.

In summary, clone YN1/1.7.4 is a standard tool for functional blockade and imaging of ICAM-1 in mouse models, uniquely enabling detailed studies of immune cell adhesion, trafficking, and inflammatory responses due to its epitope specificity, functional potency, and proven utility in diverse scientific assays.

Dosing regimens of clone YN1/1.7.4 (anti-mouse CD54/ICAM-1) show considerable variation depending on the mouse model, experimental purpose, administration route, and target tissue.

Key findings from the literature include:

• General Variability: Dose amount, dosing schedule, and administration method for clone YN1/1.7.4 are adjusted based on study goals and experimental design. There is no single, universally accepted regimen; protocols should be optimized for each application.

• In Vivo Imaging – Systemic Administration: In a study investigating local cell dynamics in vivo, mice received intraperitoneal injections of 100 μg per mouse (clone YN1/1.7.4) three times per week for two weeks prior to analysis. This regimen is typical for in vivo blockade/labeling of ICAM-1.

• Fluorescence Imaging – Local Perfusion: In confocal laser endomicroscopy of isolated liver segments, segmental perfusion doses ranged from 200 ng to 2,400 ng:

  • Minimal effective local dose: 800–1,200 ng per perfused segment.
  • Lower and higher doses: Capture efficacy was similar at 200 and 1,200 ng; however, capture efficacy significantly decreased at a high dose (2,400 ng).
  • For anti-CD54 (YN1/1.7.4), the local tissue concentration reached approximately 1.8 μg/g tissue using a 1,200-ng dose.

• Flow Cytometry & Western Blots (in vitro/ex vivo):

  • Flow cytometry: Typical concentrations are ≤0.25 μg per 1 million cells in 100 μL.
  • Western blot: 1–10 μg/mL is typical, with titration recommended.

Summary Table: Reported Dosing Regimens for Clone YN1/1.7.4

Important considerations:

• Optimization is essential: Investigators should titrate clone YN1/1.7.4 for their specific experimental system, strain, and route, as dose-responses and tissue-specific delivery may vary significantly.
• Administration method matters: Systemic (e.g., intraperitoneal) versus local (e.g., perfusion) delivery produces different tissue exposure levels and requires independent optimization.
• Tissue specificity: Capture and efficacy can depend on tissue type and perfusion; higher local doses do not always improve target engagement.

There is no universal protocol, and all studies stress the importance of experimental titration and pilot dose-response analysis for new models/applications.