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

Clone 3A6 is used in in vivo mouse studies primarily for the detection and characterization of enteroviruses (EVs), especially Coxsackievirus B (CVB) in infection models, and as an anti-Ebola virus therapeutic in some contexts.

Key in vivo applications in mice:

• Detection of Enterovirus Infections:
  Clone 3A6, originally a rat monoclonal antibody against CVB1 VP1 protein, is validated for in vivo use in mice to detect a broad range of enteroviruses. It shows strong reactivity across multiple CVB serotypes and is used to study the presence, localization, and morphology of enterovirus infections, especially in pancreatic, cardiac, and other tissues in mouse models.

• Immunohistochemistry and Immunofluorescence:
  3A6 is frequently used for immunohistochemistry (IHC) and immunofluorescence (IFA) on paraffin-embedded or PFA-fixed tissue sections from infected mice. It helps visualize viral proteins and, when combined with other markers (like dsRNA with the J2 antibody), allows co-localization studies of viral replication and protein expression within infected cells.

• No Cross-Reactivity with Mouse Proteins:
  3A6 is advantageous in mouse models because, as a rat antibody, it shows no significant cross-reactivity to mouse cellular proteins. This specificity reduces background and increases reliability in infection studies.

• Validation for Diagnostic Use:
  3A6 is described as a valid diagnostic tool in murine infection models and has been used for quality control in culturing and purifying enteroviruses for research.

• Therapeutic and Viral Targeting Research:
  Beyond detection, clone 3A6 has also been applied in studies as a therapeutic candidate (notably as an anti-Ebola virus agent) in murine infection models. However, its primary and best-established use in vivo is for enterovirus research.

Other potential in vivo applications in mice (less common):

• While clone 3A6 was originally raised against human CD166 (ALCAM), most references for its in vivo use in mice relate to viral detection rather than immunomodulation or tumor targeting, suggesting limited application in those areas unless cross-reactivity or a humanized mouse model is involved.

• Commercial sources highlight its use in functional in vivo assays, but the published literature supports its primary role in diagnostic and research applications related to viral infection.

Summary Table: 3A6 in vivo applications in mice

No significant reports indicate clone 3A6 is commonly used for targeting murine ALCAM (CD166) in vivo, as it is highly specific for the human antigen. It is possible that 3A6 is used in mouse models engrafted with human tissues, but this is not its primary in vivo application in standard mouse research.

The antibody 3A6 is used in a variety of contexts targeting different antigens (notably, Ebola virus GP, Coxsackievirus B1 VP1, CD166/ALCAM, IL-1β, and MIA), so the choice of commonly used co-antibodies or proteins depends on the research application. The answer below references the most well-established contexts from the literature:

1. 3A6 for Enterovirus Capsid Protein VP1 (Rat mAb, anti-CVB1/VP1)

• Commonly used co-antibodies/proteins:

  • Mouse-derived antibodies against enteroviral VP1 for double-staining, to enable discrimination by isotype and species.
  • Markers for tissue histology (e.g., anti-insulin or anti-glucagon for pancreatic islets, anti-trypsin for exocrine pancreas) to identify infected cell types in tissue sections.
  • Secondary antibodies: Anti-rat IgG (when used with mouse and rabbit primary antibodies).

  Relevant context: 3A6’s rat origin is specifically highlighted as enabling dual or triple labeling with mouse/rabbit antibodies in immunohistochemistry and immunofluorescence assays on mouse and human tissue samples.

2. 3A6 for Ebola Virus Glycoprotein (Human mAb, anti-EBOV GP)

• Commonly used co-antibodies/proteins:

  • Other anti-Ebola virus mAbs, such as:
    • 1A2: Targets the EBOV GP2 fusion loop.
    • 7G7: Binds to a different epitope on EBOV GP1,2.
    • Anti-influenza A (FLUAV) mAb 42–2D2: Used as an unrelated isotype control in therapy studies.
  • In neutralization panels or therapeutic cocktails, anti-GP antibodies of different specificities (e.g., REGN-EB3/ZYB3 cocktail).

  Multiple studies combine 3A6 with these anti-EBOV antibodies to compare efficacy or coverage in therapeutic models.

3. 3A6 for CD166/ALCAM (Mouse mAb, anti-human CD166/ALCAM)

• Commonly used co-antibodies/proteins:
  • Cell surface markers: Anti-CD44, anti-CD24, anti-EpCAM—used in flow cytometry to analyze stem cell or cancer cell populations alongside CD166.
  • Other adhesion molecule or differentiation markers in multiparametric cell sorting panels.

4. 3A6 for IL-1β (Mouse mAb)

• Commonly used co-antibodies/proteins:
  • Antibodies against other cytokines: e.g., anti-IL-6, anti-TNFα, for multi-cytokine detection in immunoassays.
  • Inflammatory cell markers: CD45, CD11b, used for phenotyping cytokine production in leukocytes.

5. 3A6 for MIA (Melanoma Inhibitory Activity) (Abnova, Human target)

• Commonly used co-antibodies/proteins:
  • Other tumor markers: Example, S100, HMB-45, and MART-1 in melanoma panels.
  • Housekeeping proteins: β-actin, GAPDH, for Western blot controls.

General Considerations

• Secondary antibodies tailored to the species and isotype of 3A6 are routinely used, especially in multiplex staining.
• Multiplex assays often pair 3A6 with markers that distinguish cell types, infection status, or allow for colocalization studies relevant to the disease model.

Summary Table: Common Co-antibodies/Proteins Used with 3A6

If you have a specific antigen (e.g., Ebola, enterovirus, CD166) or application in mind, I can tailor this further to that context.

The key findings from scientific literature citing clone 3A6—specifically referencing the anti-Ebola virus monoclonal antibody (mAb) 3A6—are as follows:

• Potent Protection Against Ebola Virus: 3A6 was shown to provide complete therapeutic protection in both guinea pig and rhesus monkey models at advanced, high-viremia stages of Ebola virus disease (EVD). Single administration in rhesus monkeys with severe disease rapidly reduced high viral loads to undetectable levels and reversed clinical signs of disease, at doses far lower than other approved mAb therapeutics.

• Epitope and Mechanism of Action: Through alanine scanning mutagenesis, the critical residues for 3A6 binding on the Ebola glycoprotein (GP) were identified, especially D632 and P636. The mutation P636S, for instance, abolished 3A6’s neutralizing activity, confirming the epitope specificity to the GP1,2 stalk–membrane proximal external region (MPER) of Ebola virus. This binding is believed to block essential conformational changes in the viral GP necessary for membrane fusion and viral entry, distinguishing it mechanistically from other antibodies.

• Therapeutic Window: The studies highlight that 3A6 may be uniquely effective for patients with highly advanced EVD and could expand the treatable window compared to existing antibody therapies, offering a new approach for intervention in critically ill individuals.

• Blueprint for Vaccine and Therapeutic Development: 3A6’s mechanism—targeting the flexible stalk-MPER on EBOV GP—provides a structural template for designing broad-spectrum filovirus vaccines or additional cross-reactive antibodies. The data suggest that stalk/MPER-targeted mAbs can elicit potent and broad immune responses and may be effective at even lower concentrations.

A few additional references mention other "3A6" clones in different contexts (e.g., anti-human CD166/ALCAM for immunology research, or anti-Lamin A/C antibodies for cell biology). However, the most significant and highly cited findings currently center on the anti-Ebola virus mAb 3A6 as summarized above.

There are no published or standardized in vivo dosing regimens for clone 3A6 (rat anti-EV VP1) across different mouse models. The documented use of 3A6 is largely limited to in vitro and ex vivo applications, and its use in mouse models has not been detailed in terms of dose, frequency, or route of administration.

Essential context and supporting details:

• Clone 3A6 has been validated as a rat monoclonal antibody useful in mouse models for immunohistochemistry (IHC), immunofluorescence (IFA), and Western blot due to the absence of cross-reactivity with mouse proteins. However, these validations have not included dosing regimens for in vivo administration.
• Key suppliers and research summaries emphasize that no published dosing schedule exists for 3A6 in any mouse model.

Practical implications:

• In the absence of published data, researchers may reference standard practices for rat monoclonal antibody dosing in mice; however, these are generalized guidelines and not specific to 3A6.
• Typical dosing for rat monoclonal antibodies (used in depletion or blockade studies) in mice ranges from 100–250 μg per mouse administered intraperitoneally every 3 days. This convention may be used as a starting point, but specific dose-response, safety, and efficacy for 3A6 must be empirically determined for each experimental context.

Conclusion:
Currently, dosing regimens for clone 3A6 in mouse models are neither published nor standardized. Any use of 3A6 in mice for in vivo applications should be carefully piloted, with dosing drawn from analogous antibodies and adjusted based on experimental need and safety observations.