Anti-Human CD166 (ALCAM) (Clone 3A6) – Purified in vivo GOLD^TM Functional Grade

Clone 3A6, in the context of different applications, has been used in various ways in mice:

1. Anti-Human CD166 (ALCAM) Antibody: The mouse monoclonal antibody clone 3A6 is directed against human CD166 (ALCAM), a cell adhesion molecule. It is often used in in vivo mouse studies, particularly for investigating immune responses and cell adhesion phenomena. However, its direct application in mice is primarily as a tool for studying human CD166, which may not be directly applicable to mouse models unless involving humanized mice or xenograft models.

2. Enterovirus Detection: Another 3A6 antibody, which is a rat monoclonal antibody, has been used in mouse models for detecting enteroviruses. This antibody is effective in detecting Coxsackievirus B1 (CVB1) and other enteroviruses, making it useful for studying viral infections in mouse models.

In summary, the common in vivo applications of clone 3A6 in mice depend on the specific context—either as an anti-CD166 antibody for studying adhesion molecules in humanized or xenograft models or as a detection tool for enteroviruses in murine models of infection.

Several different antibodies or proteins are commonly co-used with 3A6 in the literature, but the answer depends on which 3A6 antibody is meant, as multiple distinct monoclonals (with the clone name 3A6) are used against different targets. The most prominent in recent literature are:

1. 3A6 anti-Ebola Virus (mAb 3A6)

• Commonly used with:
  • 1A2 IgG (targets the Ebola GP2 fusion loop)
  • 7G7 IgG (targets a separate, unknown epitope on EBOV GP1,2)
  • Anti-influenza A virus antibody 42–2D2 (used as a negative control)
• These antibodies are used together in comparative protection and neutralization assays to benchmark the efficacy of 3A6 and to study combinations that may offer synergistic or broader protection.

2. 3A6 anti-CD166/ALCAM (monoclonal used in human cell adhesion studies)

• Commonly used with:
  • Other lineage markers for flow cytometry, such as:
    • CD45 (pan-leukocyte marker)
    • CD34 (hematopoietic progenitor marker)
    • CD44, CD90 (other stem cell or adhesion molecules)
    • CD6 (physiological ligand for CD166)
  • Used in panels to characterize various immune and stromal cell populations, or for co-staining in tissue sections and immunophenotyping.
• In functional studies, may be combined with antibodies against ligands or other adhesion molecules (e.g., CD6, CD54).

3. 3A6 anti-CVB1-VP1 (enterovirus detection)

• Commonly used with:
  • Pan-enterovirus antibodies or mouse-derived enteroviral capsid antibodies, since 3A6 is rat-derived and enables multiplexing.
  • Virus-specific antibodies: to confirm broad-spectrum detection across Coxsackievirus B types and Poliovirus 3.
• Used as part of immunohistochemistry or ELISA panels for enterovirus detection or double staining.

Summary Table

The specific antibody combinations depend on the target and experimental objective:

• In Ebola studies, other anti-GP antibodies are most common comparators.
• In cell marker research, other lineage and differentiation markers are routinely co-used.
• In enterovirus diagnostics, pan-EV and serotype-specific antibodies are commonly paired.

If you have a particular field or assay in mind, providing more context could narrow the list further.

Clone 3A6 has been cited in scientific literature primarily for its role as a monoclonal antibody with significant therapeutic potential against Ebola virus disease (EVD) and as a diagnostic tool in oncology.

Key findings from scientific literature include:

• Potent therapeutic effect against Ebola virus: mAb 3A6 demonstrated complete post-exposure protection in advanced EVD animal models, especially in rhesus monkeys and guinea pigs with high viral loads and late-stage disease. A single dose reduced viremia from (10^9)–(10^{10}) PFU/mL to undetectable levels by day 21, reversing clinical signs and normalizing biochemical indicators. 3A6's effective dose was significantly lower than that required for other monoclonal therapies, such as mAb114 or the REGN-EB3 cocktail.

• Unique mechanism of action: 3A6 binds to the stalk–membrane proximal external region (MPER) of the Ebola glycoprotein GP1,2. This binding is distinct from other neutralizing antibodies targeting the glycan cap or base regions and is hypothesized to block key conformational changes necessary for viral fusion with host cell membranes, thereby preventing infection progression.

• Epitope specificity: Alanine-scanning mutagenesis identified critical binding residues (such as D632A and P636A) on GP1,2; mutations at P636 (to S or Q) specifically disrupted 3A6 binding and neutralization, illustrating the antibody's precise epitope recognition and the risk of resistance if the virus mutates at these sites.

• Diagnostic and research utility in oncology: The 44-3A6 monoclonal antibody variant was generated against antigens from the A549 human lung adenocarcinoma cell line. It recognizes a cell surface antigen (40 kDa, protease-sensitive, not shed in serum or culture supernatant), providing selective detection for lung carcinomas with "glandular" differentiation. Its specificity has potential clinical and diagnostic value.

• Immunological research applications: Other variants of 3A6 target human CD166 (ALCAM), which is expressed on a broad range of cell types, including immune and cancer stem cells, and is used for cell adhesion, immunology, and neuroscience research.

Implications and future directions:

• The exceptional potency of 3A6, especially at low doses and late disease stages, could expand therapeutic options for advanced EVD cases, where current antibody treatments are less effective.
• Structural studies inform vaccine design, illustrating the importance of targeting flexible stalk–MPER regions to elicit broad and potent immune responses.
• In cancer diagnostics, 3A6-derivatives remain valuable for distinguishing specific tumor types and cell populations.

Summary of recent impact:
Clone 3A6 is a leading antibody for therapeutic, diagnostic, and fundamental research settings, with its Ebola-specific variant being noteworthy for revolutionizing late-stage disease interventions and inspiring next-generation antibodies with broader viral spectrum activity.

There are no published or standardized dosing regimens for clone 3A6 in different mouse models. The scientific literature and product datasheets confirm that 3A6, a rat monoclonal antibody (often anti-EV VP1, and sometimes referenced as anti-human CD166), lacks specific in vivo dose schedules established through published research for mouse studies.

Essential context:

• Analogous dosing approach: In the absence of 3A6-specific data, dosing regimens for other rat monoclonal antibodies used in mice can serve as a guideline. Standard practice involves administering 100–300 μg per mouse either intraperitoneally or intravenously every 3 days as an initial regimen.
• This approach is based on commonly used doses for rat monoclonal antibodies in mouse in vivo studies and is not specific to the antigen or clone, but rather a general starting point.

Supporting details:

• Validation in mouse models: 3A6 has been validated for use in mouse models for in vitro and in vivo applications, mainly immunohistochemistry and immunofluorescence, but without reporting specific dosing regimens for functional in vivo studies such as depletion or blockade.
• No cross-reactivity concern: 3A6 is advantageous in mice due to the lack of cross-reactivity with mouse proteins, reducing background issues in immunoassays, but this does not impact dosing.

Additional relevant information:

• The lack of a published protocol means that initial dosing would need to be empirically optimized for the specific experimental context, starting with the general range used for rat IgG in mice.
• General immunology protocols recommend doses for functional antibodies (such as depletion or agonist antibodies) typically within the 50–300 μg per mouse range, administered 1–3 times per week, which aligns with the suggested starting point.

Summary table of available information:

There is currently no evidence in the literature of model-specific variations for 3A6 dosing; all guidance is based on analogy to similar rat monoclonal antibodies used in mice.