ALCAM/CD166 Antibody

R&D Systems | Catalog # AF1172

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Canine

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunofluorescence, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all ALCAM/CD166 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse ALCAM/CD166
Trp28-Lys527
Accession # AAC06342

Specificity

Detects human, mouse and rat ALCAM/CD166 in Western blots. In direct ELISAs, less than 10% cross-reactivity with rhBCAM, recombinant mouse (rm) OCAM, and rmMAdCAM‑1 is observed. Detects canine ALCAM/CD166 in flow cytometry.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for ALCAM/CD166 Antibody

Detection of Human, Mouse, and Rat ALCAM/CD166 antibody by Western Blot.

Detection of Human, Mouse, and Rat ALCAM/CD166 by Western Blot.

Western blot shows lysates of mouse brain (cerebellumn) tissue, rat brain (cerebellum) tissue, H4-II-E-C3 rat hepatoma cell line, and U-118-MG human glioblastoma/astrocytoma cell line. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Mouse/Rat/Canine ALCAM/CD166 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1172) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for ALCAM/CD166 at approximately 90-120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of ALCAM/CD166 antibody in Mouse Splenocytes antibody by Flow Cytometry.

Detection of ALCAM/CD166 in Mouse Splenocytes by Flow Cytometry.

Mouse splenocytes either (A) activated or (B) resting were stained with Goat Anti-Mouse/Rat/Canine ALCAM/CD166 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1172) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107) and Rat Anti-Mouse B220/CD45R APC-conjugated Monoclonal Antibody (Catalog # FAB1217A). Quadrant markers were set based on control antibody staining (Catalog # AB-108-C).

Detection of ALCAM/CD166 in PBMC by Flow Cytometry

Human PBMC's were stained with Mouse Anti-Human CD3 epsilon PE‑conjugated Monoclonal Antibody (Catalog # FAB100P) and either (A) Goat Anti-Human/Mouse/Rat/Canine ALCAM/CD166 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1172) or (B) isotype control antibody (Catalog # AB-108-C) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). View our protocol for Staining Membrane-associated Proteins.
Detection of ALCAM/CD166 antibody in Canine PBMCs antibody by Flow Cytometry.

Detection of ALCAM/CD166 in Canine PBMCs by Flow Cytometry.

Canine peripheral blood mononuclear cells (PBMCs) treated with PMA and Calcium Ionomycin for 24 hours were stained with Goat Anti-Mouse/Rat/Canine ALCAM/CD166 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1172, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).
ALCAM/CD166 antibody in Mouse Liver by Immunohistochemistry (IHC-Fr).

ALCAM/CD166 in Mouse Liver.

ALCAM/CD166 was detected in perfusion fixed frozen sections of mouse liver using Goat Anti-Mouse/Rat/Canine ALCAM/CD166 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1172) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
ALCAM/CD166 antibody in Rat Mesenchymal Stem Cells by Immunocytochemistry (ICC).

ALCAM/CD166 in Rat Mesenchymal Stem Cells.

ALCAM/CD166 was detected in immersion fixed undifferentiated rat mesenchymal stem cells using Goat Anti-Mouse/Rat/Canine ALCAM/CD166 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1172) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Mouse ALCAM/CD166 antibody by Simple WesternTM.

Detection of Mouse ALCAM/CD166 by Simple WesternTM.

Simple Western lane view shows lysates of mouse brain tissue and mouse lung tissue loaded at 0.2 mg/mL. A specific band was detected for ALCAM/CD166 at approximately 149 kDa (as indicated) using 12.5 µg/mL of Goat Anti-Mouse/Rat/Canine ALCAM/CD166 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1172) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for ALCAM/CD166 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Mouse splenocytes, canine peripheral blood mononuclear cells (PBMCs) treated with PMA and Calcium Ionomycin or CD3+ Human PBMC's.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed undifferentiated rat mesenchymal stem cells

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of mouse liver and thymus

Simple Western

12.5 µg/mL
Sample:   Mouse brain (total) tissue and mouse lung tissue

Western Blot

0.2 µg/mL
Sample: Mouse brain (cerebellumn) tissue, rat brain (cerebellum) tissue, H4‑II‑E‑C3 rat hepatoma cell line, and U‑118‑MG human glioblastoma/astrocytoma cell line

Reviewed Applications

Read 2 reviews rated 4.5 using AF1172 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ALCAM/CD166

ALCAM, activated leukocyte cell adhesion molecule, is a type I membrane glycoprotein and a member of the immunoglobulin supergene family. It is also known as CD166, MEMD, SC-1/DM-GRASP/BEN in the chicken, and KG-CAM in the rat. ALCAM is expressed on thymic epithelial cells, activated B and T cells, and monocytes. ALCAM can bind itself homotypically and is also capable of binding CD6, NgCAM, and other, as of yet, unidentified brain proteins. ALCAM/CD6 interaction may be involved in T cell development and T cell regulation. Additionally, ALCAM/CD6 and ALCAM/NgCAM interactions may play roles in the nervous system. ALCAM has also been observed to be upregulated on highly metastasizing melanoma cell lines and may play a role in tumor migration. ALCAM is a 583 amino acid (aa) protein consisting of a 27 aa signal peptide, a 500 aa extracellular domain, a 24 aa transmembrane domain, and a 32 aa cytoplasmic domain. The extracellular domain of ALCAM contains 5 Ig-like domains of which the amino-terminal V1 domain is essential for ligand binding and ALCAM-mediated cell aggregation (1-4).  The ECD of mouse ALCAM shares 97.5% and 93.2% aa sequence identity with rat and canine ALCAM ECD, respectively.

References

  1. Bowen, M.A. et al. (1995) J. Exp. Med. 181:2213.
  2. Aruffo, A. et al. (1997) Immunol. Today 18:498.
  3. Degen, W.G. et al. (1998) Am. J. Pathol. 152:805.
  4. Van Kempen, L. et al. (2001) J. Biol. Chem. 276:25783.

Long Name

Activated Leukocyte Cell Adhesion Molecule

Alternate Names

CD166

Entrez Gene IDs

214 (Human); 11658 (Mouse); 101867493 (Cynomolgus Monkey)

Gene Symbol

ALCAM

UniProt

AAC06342

Additional ALCAM/CD166 Products

Product Documents for ALCAM/CD166 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for ALCAM/CD166 Antibody

For research use only

Citations for ALCAM/CD166 Antibody

Customer Reviews for ALCAM/CD166 Antibody (2)

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  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 23169771
    Species: Mouse
    Verified Customer | Posted 01/05/2015
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: See PMID 22243278
    Species: Mouse
    Verified Customer | Posted 01/05/2015

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