Anti-Human CD166 (ALCAM) [Clone 3A6] — Purified in vivo PLATINUM™ Functional Grade

The monoclonal antibody clone 3A6 has been used in in vivo mouse studies primarily for two distinct applications depending on antigen specificity and antibody origin:

1. Ebola Virus (EBOV) Monoclonal Antibody 3A6

• Prophylactic and Therapeutic Use: Mouse studies show that prophylactic administration of 3A6 IgG provides robust protection against lethal challenge with mouse-adapted Ebola virus. A dose of 100??g (about 5?mg/kg) gave complete (100%) protection, while 25??g (1.25?mg/kg) provided partial (50%) protection in mice.
• Notable Efficacy in Stringent Models: 3A6 has also demonstrated strong post-exposure efficacy in guinea pigs and rhesus monkeys challenged with Ebola, suggesting broad in vivo potency.
• Mode of Use: Antibodies are typically administered intravenously or intraperitoneally for these experiments.

2. Rat-derived 3A6 Against Coxsackievirus (CVB1-VP1) and Enteroviruses

• Detection in Mouse Tissues: The rat monoclonal antibody clone 3A6 targets a group-specific epitope on the VP1 protein of enteroviruses. It has been validated for detection of acute Coxsackievirus and other enteroviral infections in murine tissues by immunohistochemistry (IHC).
• Advantage in Mouse Models: As a rat antibody, 3A6 is particularly advantageous for use in mouse studies, allowing clear immunohistochemical detection of viral antigens without problematic background from mouse endogenous antibodies.
• Applications: Used to stain infected murine tissues (e.g., pancreas) to reveal viral localization and tissue tropism during enterovirus infection, with specificity for EV-Bs and key EV-Cs.

Clarification

There are multiple monoclonal antibodies named "3A6" targeting different proteins. Based on context, in in vivo mouse studies:

• Ebola virus 3A6 is used for protection and therapeutic studies after viral challenge.
• EV-VP1 rat 3A6 is used for viral detection in mouse tissue sections without cross-reactivity to mouse proteins, key for enterovirus research.

If you are referring to another 3A6 (e.g., anti-human CD166/ALCAM), please specify, as each is distinct in application and target.

The correct storage temperature for most sterile packaged laboratory reagents or biological materials, including monoclonal antibody clones like 3A6, is typically specified by the manufacturer, but commonly falls within the 2°C to 8°C range for long-term preservation of sterility and function.

Essential details:

• For biologicals such as vaccines and certain reagents, storage in a refrigerator between 2°C and 8°C (35°F to 46°F) is standard to maintain sterility and efficacy.
• The packaging or product insert from the manufacturer should always be checked for specific storage requirements for clone 3A6, as deviations (such as freezing or temperatures above 8°C) may damage the viability or performance of the reagent.
• If the product is not a biological reagent but rather medical equipment or sterile devices, ambient sterile storage requirements apply: Controlled environments with temperatures up to 24°C (75°F) and relative humidity between 30-60% are often recommended to prevent microbial growth.

If your "clone 3A6" is a sterile packaged monoclonal antibody or cell line, store it at 2°C–8°C unless the manufacturer's datasheet or label states otherwise. Do not freeze unless instructed by the manufacturer. Always protect the product from light if instructed.

If you have access to the product datasheet or sterile package, follow its specific recommendations for optimal storage.

The most commonly co-used antibodies with 3A6 in the literature depend on the specific 3A6 antibody and its application, as multiple antibodies called "3A6" exist for different targets:

• For Coxsackievirus B1 VP1 (rat monoclonal 3A6):
  The literature often pairs 3A6 with 5D8/1, a widely used mouse monoclonal antibody against enterovirus VP1.

  • 3A6 offers the advantage of a different host species (rat), enabling its use in double staining protocols together with 5D8/1 in both human and mouse tissue samples without cross-reactivity issues.
  • 5D8/1 has also been noted to show some non-specific reactivity (e.g., to smooth muscle), whereas 3A6 does not.
  • Frequently, 3A6 is used in conjunction with generic detection of enteroviral proteins or with cellular/tissue markers appropriate to the studied system for localization and co-localization studies.

• For Ebola Virus GP1,2 (human mAb 3A6):
  Pairings commonly involve assessing 3A6 in tandem or comparison with other anti-GP1,2 antibodies such as BDBV223 or antibodies targeting different GP1,2 epitopes.

  • In mechanistic or escape studies, BDBV223 is often used as a point of comparison since both recognize the HR2 stalk–MPER region.
  • Supporting detection secondary antibodies such as DyLight 488 anti-human IgG are employed for visualization in immunofluorescence and flow cytometry.
  • Other anti-fusogenic or MPER-targeted antibodies (e.g., those for HIV GP160 or influenza hemagglutinin) may be mentioned in mechanistic studies for comparison, though not always used concurrently in assays.

• For CD166/ALCAM (mouse mAb 3A6):
  This 3A6 is used in flow cytometric panels alongside lineage markers or disease-relevant markers to identify or characterize cell populations.

  • Typically, co-staining involves pan-leukocyte markers (e.g., CD45), stem/progenitor cell markers, or other adhesion molecules, depending on the experiment.

Key points:

• When 3A6 (rat anti-CVB1-VP1) is used, 5D8/1 (mouse anti-VP1) is the most frequent co-applied antibody.
• When 3A6 (anti-Ebola GP1,2) is used, BDBV223 and similar anti-GP1,2 antibodies are used for comparison or combination.
• When 3A6 (anti-CD166/ALCAM) is used, typical flow cytometry panels apply it with other cell surface marker antibodies for multiparametric analysis.

Each 3A6 clone is target-specific, so the choice of accompanying antibodies depends entirely on the antigen and experimental context. If you specify which 3A6 you mean, more tailored details can be provided.

Key findings regarding "clone 3A6" in scientific literature refer to several independent research tools and antibodies named "3A6," each with distinct targets and applications. The most notable are:

1. 3A6 as a Rat Monoclonal Antibody Against Enteroviruses (EVs)

• Broad reactivity: The rat monoclonal antibody clone 3A6 recognizes a group-specific epitope in the N-terminus of the enterovirus VP1 protein, including several EV-Bs and Poliovirus 3 (EV-C), making it a broad-spectrum EV-specific antibody.
• Versatility: 3A6 performs well in multiple assay platforms—including ELISA, immunoelectron microscopy, immunocytochemistry, immunohistochemistry, and Western blotting—enabling detection of enteroviruses in infected cells and tissues for both research and diagnostic applications.
• Species advantage: As a rat-derived antibody, 3A6 circumvents the high background and specificity problems associated with commonly used mouse monoclonal antibodies in murine samples, improving double-staining protocols and utility in mouse models.

2. 3A6 Monoclonal Antibody against Ebola Virus (EBOV)

• Potent therapeutic efficacy: In animal models (guinea pig and rhesus monkey), 3A6 provided complete post-exposure protection against advanced lethal Ebola virus disease, even at high viremia levels, at lower doses compared to existing antibody therapies.
• Novel mechanism: 3A6 binds a specific epitope in the stalk–membrane-proximal external region (MPER) of the Ebola glycoprotein (GP1,2), likely blocking conformational changes required for virus–cell membrane fusion.
• Resistance and specificity: Escape mutations (e.g., P636S in GP1,2) eliminate 3A6 binding and neutralization capacity, highlighting epitope specificity and the necessity of this residue for activity.
• Therapeutic window: The unparalleled potency and activity at advanced disease stages suggest 3A6 may extend the therapeutic window for Ebola virus infection beyond current therapies.

3. Human Autoimmune T-cell Clone 3A6 (TCR studies in MS)

• Multiple sclerosis model: Human CD4+ T-cell clone 3A6 recognizes the immunodominant myelin basic protein (MBP) 84–102 epitope in HLA-DR2a+ individuals, including MS patients; transgenic expression of 3A6 TCR in mice induces central nervous system autoimmunity mimicking MS.
• Antigen recognition: 3A6 TCR demonstrates highly degenerate peptide recognition, tolerating substitutions at most TCR–contacting sites and indicating broad autoreactivity potential.
• Pathogenic relevance: In vivo, 3A6 recapitulates disease features, enabling mechanistic studies of antigen specificity and pathogenesis in autoimmune demyelinating disease.

Summary Table – Distinct 3A6 Clones in Scientific Use

Each 3A6 clone has made significant contributions to virology, immunology, diagnostics, and therapeutic research. The precise impact depends on field of application and target epitope. If you need findings about a specific 3A6 context, please clarify which (e.g., enterovirus antibody, Ebola antibody, autoimmune TCR).