Human/Mouse/Rat/Canine ALCAM/CD166 Antibody

Catalog # Availability Size / Price Qty
AF1172
AF1172-SP
Detection of Human, Mouse, and Rat ALCAM/CD166 by Western Blot.
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Product Details
Citations (22)
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Human/Mouse/Rat/Canine ALCAM/CD166 Antibody Summary

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 either lyophilized or 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.

Scientific Data

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

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.

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

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).

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

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 antibody in Mouse Liver by Immunohistochemistry (IHC-Fr). View Larger

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 antibody in Rat Mesenchymal Stem Cells by Immunocytochemistry (ICC). View Larger

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.

Simple Western Detection of Mouse ALCAM/CD166 antibody by Simple Western<sup>TM</sup>. View Larger

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.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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.

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); 101867493 (Cynomolgus Monkey)
Alternate Names
activated leucocyte cell adhesion molecule; activated leukocyte cell adhesion moleculeFLJ38514; ALCAM; CD166 antigen; CD166; MEMDMGC71733

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Citations for Human/Mouse/Rat/Canine ALCAM/CD166 Antibody

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.

22 Citations: Showing 1 - 10
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  1. Chromatin accessibility dynamics of neurogenic niche cells reveal defects in neural stem cell adhesion and migration during aging
    Authors: Robin W. Yeo, Olivia Y. Zhou, Brian L. Zhong, Eric D. Sun, Paloma Navarro Negredo, Surag Nair et al.
    Nature Aging
  2. VE-cadherin in arachnoid and pia mater cells serves as a suitable landmark for in vivo imaging of CNS immune surveillance and inflammation
    Authors: Mapunda JA, Pareja J, Vladymyrov M et al.
    Nat Commun
  3. CD166-specific CAR-T cells potently target colorectal cancer cells
    Authors: S He, S Li, J Guo, X Zeng, D Liang, Y Zhu, Y Li, D Yang, X Zhao
    Translational Oncology, 2022-10-31;27(0):101575.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: Flow Cytometry, ICC, Western Blot
  4. Neural cell adhesion molecule is required for ventricular conduction system development
    Authors: Camila Delgado, Lei Bu, Jie Zhang, Fang-Yu Liu, Joseph Sall, Feng-Xia Liang et al.
    Development
  5. A comprehensive library of human transcription factors for cell fate engineering
    Authors: AHM Ng, P Khoshakhla, JE Rojo Arias, G Pasquini, K Wang, A Swiersy, SL Shipman, E Appleton, K Kiaee, RE Kohman, A Vernet, M Dysart, K Leeper, W Saylor, JY Huang, A Graveline, J Taipale, DE Hill, M Vidal, JM Melero-Mar, V Busskamp, GM Church
    Nature Biotechnology, 2020-11-30;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  6. Endophilin-A3 and Galectin-8 control the clathrin-independent endocytosis of CD166
    Authors: HF Renard, F Tyckaert, C Lo Giudice, T Hirsch, CA Valades-Cr, C Lemaigre, M Shafaq-Zad, C Wunder, R Wattiez, L Johannes, P van der Br, D Alsteens, P Morsomme
    Nat Commun, 2020-03-19;11(1):1457.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: IF
  7. Hepatocyte-like Cells Derived from Human Pluripotent Stem Cells Can Be Enriched by a Combination of Mitochondrial Content and Activated Leukocyte Cell Adhesion Molecule
    Authors: Hiromi Yamashita, Keiichi Fukuda, and Fumiyuki Hattori
    Japan Medical Association Journal
    Species: Human, Mouse
    Sample Types: Whole Cells
    Applications: IHC, Flow Cytometry
  8. Axonal growth of midbrain dopamine neurons is modulated by the cell adhesion molecule ALCAM through trans-heterophilic interaction with L1Cam, ChL1 and Semaphorin
    Authors: CR Bye, V Rytova, WF Alsanie, CL Parish, LH Thompson
    J. Neurosci., 2019-07-12;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  9. ALCAM Mediates DC Migration Through Afferent Lymphatics and Promotes Allospecific Immune Reactions
    Authors: Ann-Helen Willrodt, Ann-Charlott Salabarria, Philipp Schineis, Desislava Ignatova, Morgan Campbell Hunter, Martina Vranova et al.
    Frontiers in Immunology
  10. Distribution and development of molecularly distinct perineuronal nets in visual thalamus
    Authors: Ubadah Sabbagh, Aboozar Monavarfeshani, Kaiwen Su, Masoud Zabet‐Moghadam, James Cole, Eric Carnival et al.
    Journal of Neurochemistry
  11. Tbr1 instructs laminar patterning of retinal ganglion cell dendrites
    Authors: J Liu, JDS Reggiani, MA Laboulaye, S Pandey, B Chen, JLR Rubenstein, A Krishnaswa, JR Sanes
    Nat. Neurosci., 2018-04-09;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  12. Multicolor quantitative confocal imaging cytometry
    Authors: DL Coutu, KD Kokkaliari, L Kunz, T Schroeder
    Nat. Methods, 2017-11-13;15(1):39-46.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  13. Cell Adhesion Molecule CD166/ALCAM Functions Within the Crypt to Orchestrate Murine Intestinal Stem Cell Homeostasis
    Authors: Nicholas R. Smith, Paige S. Davies, Trevor G. Levin, Alexandra C. Gallagher, Douglas R. Keene, Sidharth K. Sengupta et al.
    Cellular and Molecular Gastroenterology and Hepatology
  14. 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, 2017-01-09;114(4):E524-E533.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  15. 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, 2015-03-31;10(3):e0121550.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  16. 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, 2015-03-09;112(15):E1946-55.
    Species: Mouse, Rat
    Sample Types: Tissue Homogenates, Whole Tissue
    Applications: Flow Cytometry, IHC
  17. 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, 2014-03-18;5(0):3494.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  18. Dbx1 triggers crucial molecular programs required for midline crossing by midbrain commissural axons.
    Authors: Inamata, Yasuyuki, Shirasaki, Ryuichi
    Development, 2014-02-19;141(6):1260-71.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  19. CD166/ALCAM Mediates Proinflammatory Effects of S100B in Delayed Type Hypersensitivity
    Authors: Rüdiger von Bauer, Dimitrios Oikonomou, Alba Sulaj, Sawsan Mohammed, Agnes Hotz-Wagenblatt, Hermann-Josef Gröne et al.
    The Journal of Immunology
  20. 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., 2011-04-20;31(16):5942-55.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  21. 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., 2008-06-18;28(25):6285-94.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  22. Inhibition of p38 MAPK signaling in chondrocyte cultures results in enhanced osteogenic differentiation of perichondral cells.
    Authors: Stanton LA, Beier F
    Exp. Cell Res., 2006-10-04;313(1):146-55.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC

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