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.
Human/Mouse/Rat/Canine ALCAM/CD166 Antibody
R&D Systems | Catalog # AF1172
Key Product Details
Validated by
Species Reactivity
Validated:
Cited:
Applications
Validated:
Cited:
Label
Antibody Source
Product Specifications
Immunogen
Trp28-Lys527
Accession # AAC06342
Specificity
Clonality
Host
Isotype
Scientific Data Images for Human/Mouse/Rat/Canine ALCAM/CD166 Antibody
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 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 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 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 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 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 Human/Mouse/Rat/Canine ALCAM/CD166 Antibody
CyTOF-ready
Flow Cytometry
Sample: Mouse splenocytes, canine peripheral blood mononuclear cells (PBMCs) treated with PMA and Calcium Ionomycin or CD3+ Human PBMC's.
Immunocytochemistry
Sample: Immersion fixed undifferentiated rat mesenchymal stem cells
Immunohistochemistry
Sample: Perfusion fixed frozen sections of mouse liver and thymus
Simple Western
Sample: Mouse brain (total) tissue and mouse lung tissue
Western Blot
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
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
Formulation, Preparation, and Storage
Purification
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Stability & Storage
- 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
Background: ALCAM/CD166
References
- Bowen, M.A. et al. (1995) J. Exp. Med. 181:2213.
- Aruffo, A. et al. (1997) Immunol. Today 18:498.
- Degen, W.G. et al. (1998) Am. J. Pathol. 152:805.
- Van Kempen, L. et al. (2001) J. Biol. Chem. 276:25783.
Long Name
Alternate Names
Gene Symbol
UniProt
Additional ALCAM/CD166 Products
Product Documents for Human/Mouse/Rat/Canine ALCAM/CD166 Antibody
Product Specific Notices for Human/Mouse/Rat/Canine ALCAM/CD166 Antibody
For research use only
Related Research Areas
Citations for Human/Mouse/Rat/Canine ALCAM/CD166 Antibody
Customer Reviews for Human/Mouse/Rat/Canine ALCAM/CD166 Antibody (2)
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Application: ImmunofluorescenceSample Tested: See PMID 23169771Species: MouseVerified Customer | Posted 01/05/2015
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Application: ImmunofluorescenceSample Tested: See PMID 22243278Species: MouseVerified Customer | Posted 01/05/2015
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars