hh Human/Mouse/Rat/Canine ALCAM/CD166 Antibody AF1172: R&D Systems

Human/Mouse/Rat/Canine ALCAM/CD166 Antibody

(8 citations)   
  • Species Reactivity
    Human, Mouse, Rat, Canine
  • 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.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant mouse ALCAM/CD166
    Trp28-Lys527
    Accession # AAC06342
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Simple Western
    12.5 µg/mL
    See below
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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 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).
Immunohistochemistry
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.

Immunocytochemistry
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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
  • Entrez Gene IDs:
    214 (Human); 11658 (Mouse)
  • Alternate Names:
    activated leucocyte cell adhesion molecule; activated leukocyte cell adhesion moleculeFLJ38514; ALCAM; CD166 antigen; CD166; MEMDMGC71733
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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Species
Applications
Sample Type
  1. Dual role of ALCAM in neuroinflammation and blood-brain barrier homeostasis
    Authors: MA L‚cuyer, O Saint-Laur, L BourbonniŠ, S Larouche, C Larochelle, L Michel, M Charabati, M Abadier, S Zandee, N Haghayegh, E Gowing, C Pittet, R Lyck, B Engelhardt, A Prat
    Proc. Natl. Acad. Sci. U.S.A, 2017;114(4):E524-E533.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: IP
  2. Transcriptome analysis reveals transmembrane targets on transplantable midbrain dopamine progenitors.
    Authors: Bye C, Jonsson M, Bjorklund A, Parish C, Thompson L
    Proc Natl Acad Sci U S A, 2015;112(15):E1946-55.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC - Not specified
  3. Expression of the immunoglobulin superfamily cell adhesion molecules in the developing spinal cord and dorsal root ganglion.
    Authors: Gu, Zirong, Imai, Fumiyasu, Kim, In Jung, Fujita, Hiroko, Katayama, Kei ichi, Mori, Kensaku, Yoshihara, Yoshihir, Yoshida, Yutaka
    PLoS ONE, 2015;10(3):e0121550.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Not Specified
  4. Ionizing irradiation induces acute haematopoietic syndrome and gastrointestinal syndrome independently in mice.
    Authors: Leibowitz B, Wei L, Zhang L, Ping X, Epperly M, Greenberger J, Cheng T, Yu J
    Nat Commun, 2014;5(0):3494.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IF
  5. Dbx1 triggers crucial molecular programs required for midline crossing by midbrain commissural axons.
    Authors: Inamata, Yasuyuki, Shirasaki, Ryuichi
    Development, 2014;141(6):1260-71.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC
  6. Cocaine hijacks sigma1 receptor to initiate induction of activated leukocyte cell adhesion molecule: implication for increased monocyte adhesion and migration in the CNS.
    Authors: Yao H, Kim K, Duan M, Hayashi T, Guo M, Morgello S, Prat A, Wang J, Su TP, Buch S
    J. Neurosci., 2011;31(16):5942-55.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  7. Molecular mechanisms controlling midline crossing by precerebellar neurons.
    Authors: Di Meglio T, Nguyen-Ba-Charvet KT, Tessier-Lavigne M, Sotelo C, Chedotal A
    J. Neurosci., 2008;28(25):6285-94.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Frozen
  8. Inhibition of p38 MAPK signaling in chondrocyte cultures results in enhanced osteogenic differentiation of perichondral cells.
    Authors: Stanton LA, Beier F
    Exp. Cell Res., 2007;313(1):146-55.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC
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