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

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

Product No.: C712

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Clone
3A6
Target
CD166
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
CD6 ligand, Activated Leukocyte Cell Adhesion Molecule (ALCAM)
Isotype
Mouse IgG1 κ
Applications
FC
,
IF
,
IF Microscopy
,
IHC
,
IP

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Select Product Size
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Antibody Details

Product Details

Reactive Species
Human
Host Species
Mouse
Recommended Dilution Buffer
Immunogen
Cultured human thymic epithelial cells
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<0.5 EU/mg as determined by the LAL method
Purity
≥98% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM<sup>TM</sup> antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Applications and Recommended Usage?
Quality Tested by Leinco
FC
Additional Applications Reported In Literature ?
IHC,
IP,
IF,
IF Microscopy
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
3A6 activity is directed against human CD166 (ALCAM) and cross-reacts with ovine tissues.
Background
Activated leukocyte cell adhesion molecule (ALCAM) is a member of the immunoglobulin superfamily and a cell surface glycoprotein1. In normal physiology, ALCAM functions in cell adhesion, is known to promote T cell activation and proliferation by interacting with CD6, and functions in angiogenesis, monocyte transmigration, leukocyte intravasation across the blood-brain barrier, hematopoiesis, neurite extension, osteogenesis, and embryonic implantation in the uterus. In cancer, ALCAM is a prognostic marker of disease progression and acts as a modulator of progression by controlling cell proliferation, adhesion, migration, and invasion.

ALCAM participates in homophilic ALCAM-ALCAM interactions as well as numerous heterophilic interactions1. Ligands include CD6, galectin-8, endophilin-A3/galectin-8, CD9, S100B, and ezrin. Additionally, SOSTDC1 is a novel ligand of ALCAM that promotes invasion and facilitates liver metastasis in colorectal cancer through activation of the Src-P13K/AKT pathways2.

ALCAM is a type I transmembrane molecule with a large glycosylated extracellular domain1. Two isoforms have been confirmed at the protein level: ALCAM-Iso1, which is the full length isoform, and ALCAM-Iso2, which lacks exon 13. ALCAM is proteolytically cleaved at its extracellular domain by the transmembrane metalloprotease ADAM17, with ALCAM-Iso2 more susceptible to cleavage.

3A6 was produced by immunizing mice with human thymic epithelial cells and then fusing spleen cells with P3X63Ag8 myeloma cells3. 3A6 cross reacts with ovine mesenchymal stromal cells from iliac crest bone marrow aspirates4.
Antigen Distribution
CD166 is expressed on neurons, activated leukocytes, hematopoietic stem cells, mesenchymal stem cells, bone marrow stromal cells, activated T cells, activated B cells, activated monocytes, thymic epithelial cells, vascular endothelial cells, fibroblasts, keratinocytes, myeloid progenitors, tumor cells, and cancer stem cells.
NCBI Gene Bank ID
Research Area
Cell Adhesion
.
Cell Biology
.
Immunology
.
Neuroscience
.
Synaptic Biology

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

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

Application AreaDetails and Notes
Enterovirus detection:Used for IHC/IFA to visualize and localize EV infections in tissues
Therapeutic studies:Tested as anti-Ebola virus agent (less common) in murine models
Diagnostics & QC:Quality control for EV culture and purification; minimal cross-reactivity
Immunohistochemistry/FluorescenceUsed on both acute and chronic infection models, with paraffin/PFA samples

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

3A6 Target/ContextCommon Co-antibodies/Proteins
Enterovirus VP1Mouse anti-VP1, anti-insulin, anti-trypsin, anti-glucagon
Ebola GP1A2, 7G7, anti-FLUAV 42–2D2, isotype/cytokine controls
CD166/ALCAMAnti-CD44, anti-CD24, anti-EpCAM, other stem/cancer markers
IL-1βAnti-IL-6, anti-TNFα, CD45, CD11b
MIAS100, HMB-45, MART-1, β-actin, GAPDH

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.

References & Citations

1. Ferragut F, Vachetta VS, Troncoso MF, et al. Cytokine Growth Factor Rev. 61:27-37. 2021.
2. Bartolomé RA, Pintado-Berninches L, Jaén M, et al. Oncogene. 39(38):6085-6098. 2020.
3. Patel DD, Fong AM, Mann KP, et al. CD166 Workshop: Tissue distribution and functional analysis of antibodies reactive for CD166, a ligand for CD6. In: Kishimoto T, editor. Leukocyte Typing IV. Oxford: Oxford University Press; 1997. P.461-464.
4. Sanjurjo-Rodríguez C, Castro-Viñuelas R, Hermida-Gómez T, et al. PLoS One. 12(1):e0171231. 2017.
5. Piazza T, Cha E, Bongarzone I, et al. J Cell Sci. 118(Pt 7):1515-1525. 2005.
6. Tondreau T, Dejeneffe M, Meuleman N, et al. BMC Genomics. 9:166. 2008.
7. Srouji S, Kizhner T, Ben David D, et al. Calcif Tissue Int. 84(2):138-145. 2009.
8. Katsube Y, Kotobuki N, Tadokoro M, et al. Gene Ther. 17(4):494-502. 2010.
9. Brune JC, Tormin A, Johansson MC, et al. Int J Cancer. 129(2):319-330. 2011.
10. Ali H, Al-Yatama MK, Abu-Farha M, et al. PLoS One. 10(4):e0122465. 2015.
11. Prins HJ, Schulten EA, Ten Bruggenkate CM, et al. Stem Cells Transl Med. 5(10):1362-1374. 2016.
12. Fridriksdottir AJ, Kim J, Villadsen R, et al. Nat Commun. 6:8786. 2015.
13. Gong B, Zheng L, Lu Z, et al. Mol Med Rep. 23(1):43. 2021.
14. Yeh SP, Chang JG, Lin CL, et al. Leukemia. 19(8):1505-1507. 2005.
15. Levesque MC, Heinly CS, Whichard LP, et al. Arthritis Rheum. 41(12):2221-2229. 1998.
16. Ishiguro F, Murakami H, Mizuno T, et al. J Thorac Oncol. 7(5):890-899. 2012.
17. Bhattacharya S, Mathew G, Ruban E, et al. J Proteome Res. 9(12):6112-6125. 2010.
Flow Cytometry
IF
IF Microscopy
IHC
Immunoprecipitation Protocol

Certificate of Analysis

Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.