ALCAM/CD166 Antibody (105902) [Phycoerythrin]

Human whole blood monocytes were stained with Mouse Anti-Human ALCAM/CD166 PE-conjugated Monoclonal Antibody (Catalog # FAB6561P, filled histogram) or isotype control antibody (IC002P, open histogram). View our protocol for Staining Membrane-associated Proteins.

Analysis of CSC marker expression in TOP-GFP cultures.(A) Representative images of the three independent single-cell-cloned CSC cultures, lentivirally transduced with TOP-GFP. Phase contrast (top) and fluorescence microscopy (bottom) for each of the cultures indicated. Bar = 90 µm. (B) Single parameter histograms for GFP intensity for each of the TOP-GFP single-cell-cloned CSC cultures with the TOP-GFPlow (10% lowest) and TOP-GFPhigh (10% highest) populations indicated. (C) Single parameter histograms for the indicated cell surface markers for each of the indicated cultures. Gray denotes TOP-GFPlow (10% lowest) and green denotes TOP-GFPhigh (10% highest) populations. (D) Density plots for CD29/CD24 and CD44/CD166 from TOP-GFPlow (gray) and TOP-GFPhigh (green) populations of each culture. Additional details for this experiment can be found at //osf.io/tfy28/.Flow cytometry gating strategy.Representative density plots of gating strategy to assess cell surface markers from TOP-GFPlow and TOP-GFPhigh populations. Forward scatter area (FSC-A) and PerCP-Cy5.5 was used to gate on viable cells (PI negative cells), followed by forward verses side scatter area (FSC-A vs SSC-A) to identify cells of interest and exclude debris, which were then analyzed by FSC-A and forward scatter width (FSC-W), and then SSC-A and side scatter width (SSC-W) to exclude doublet cells. From the single-cell population, SSC and FITC were used to gate on the TOP-GFPlow (10% lowest) and TOP-GFPhigh (10% highest) populations. TOP-GFPlow and TOP-GFPhigh populations were then assessed for PE and APC to detect the fluorophores conjugated to antibodies against the cell surface markers analyzed in this study. Additional details for this experiment can be found at //osf.io/tfy28/. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31215867), licensed under a CC-BY license. Not internally tested by R&D Systems.

Analysis of CSC marker expression in TOP-GFP cultures.(A) Representative images of the three independent single-cell-cloned CSC cultures, lentivirally transduced with TOP-GFP. Phase contrast (top) and fluorescence microscopy (bottom) for each of the cultures indicated. Bar = 90 µm. (B) Single parameter histograms for GFP intensity for each of the TOP-GFP single-cell-cloned CSC cultures with the TOP-GFPlow (10% lowest) and TOP-GFPhigh (10% highest) populations indicated. (C) Single parameter histograms for the indicated cell surface markers for each of the indicated cultures. Gray denotes TOP-GFPlow (10% lowest) and green denotes TOP-GFPhigh (10% highest) populations. (D) Density plots for CD29/CD24 and CD44/CD166 from TOP-GFPlow (gray) and TOP-GFPhigh (green) populations of each culture. Additional details for this experiment can be found at //osf.io/tfy28/.Flow cytometry gating strategy.Representative density plots of gating strategy to assess cell surface markers from TOP-GFPlow and TOP-GFPhigh populations. Forward scatter area (FSC-A) and PerCP-Cy5.5 was used to gate on viable cells (PI negative cells), followed by forward verses side scatter area (FSC-A vs SSC-A) to identify cells of interest and exclude debris, which were then analyzed by FSC-A and forward scatter width (FSC-W), and then SSC-A and side scatter width (SSC-W) to exclude doublet cells. From the single-cell population, SSC and FITC were used to gate on the TOP-GFPlow (10% lowest) and TOP-GFPhigh (10% highest) populations. TOP-GFPlow and TOP-GFPhigh populations were then assessed for PE and APC to detect the fluorophores conjugated to antibodies against the cell surface markers analyzed in this study. Additional details for this experiment can be found at //osf.io/tfy28/. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31215867), licensed under a CC-BY license. Not internally tested by R&D Systems.

In vitro maintenance and expansion of CoCSC.Human ESA+CD44+CD166+ cells were plated in limiting dilution and cultured for fourteen days in serum-free maintenance conditions. Colorectal tumor colonies were then either analyzed for A) ESA expression by IHC, or B) ESA, CD44 and CD166 expression by flow cytometry. C) Cellular phenotype of single colony-derived tumors, showing human ESA+ cell subpopulations expressing CD44 and CD166. D) Tumor growth curves are shown for either 2,000 CoCSC phenotype cells or an equal number of cells with all other phenotypes (Other), which were isolated from in vitro colony-derived tumors (tumors/animals injected). Inset shows lentiviral insertion band obtained by inverse PCR of 1) human xenograft tumor cells, 2) ESA+CD44+CD166+ (CoCSC) cells or 3) ESA+CD44− (Other) cells isolated by FACS. Phenotypic and morphological analysis of single-cell derived tumors from serially transplanted CoCSC show that the diverse E) phenotype and F) histological makeup of xenogeneic colorectal tumors are maintained following brief in vitro culture in limiting dilution. Black bar = 100 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/18560594), licensed under a CC-BY license. Not internally tested by R&D Systems.

In vitro maintenance and expansion of CoCSC.Human ESA+CD44+CD166+ cells were plated in limiting dilution and cultured for fourteen days in serum-free maintenance conditions. Colorectal tumor colonies were then either analyzed for A) ESA expression by IHC, or B) ESA, CD44 and CD166 expression by flow cytometry. C) Cellular phenotype of single colony-derived tumors, showing human ESA+ cell subpopulations expressing CD44 and CD166. D) Tumor growth curves are shown for either 2,000 CoCSC phenotype cells or an equal number of cells with all other phenotypes (Other), which were isolated from in vitro colony-derived tumors (tumors/animals injected). Inset shows lentiviral insertion band obtained by inverse PCR of 1) human xenograft tumor cells, 2) ESA+CD44+CD166+ (CoCSC) cells or 3) ESA+CD44− (Other) cells isolated by FACS. Phenotypic and morphological analysis of single-cell derived tumors from serially transplanted CoCSC show that the diverse E) phenotype and F) histological makeup of xenogeneic colorectal tumors are maintained following brief in vitro culture in limiting dilution. Black bar = 100 µm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/18560594), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: Flow
• Clone: 105902
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Phycoerythrin

ALCAM/CD166 Antibody (105902) [Phycoerythrin] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human ALCAM/CD166
  Trp28-Ala526
  Accession # Q13740
• Specificity: Detects human ALCAM in Western blots. Shows approximately 50% cross-reactivity with recombinant mouse OCAM and no cross-reactivity with recombinant human (rh) BCAM, rhEpCAM, rhMCAM, or rhNCAM-L1.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: ALCAM
• Purity Statement: Protein A or G purified from ascites

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 10 uL/10^6 cells

Packaging, Storage & Formulations

• Storage: Store the unopened product at 2 - 8° C. Do not use past expiration date. Protect from light.
• Buffer: Supplied in a saline solution containing BSA and Sodium Azide.
• Preservative: Sodium Azide

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ALCAM/CD166 Antibody (105902) [Phycoerythrin]

• activated leucocyte cell adhesion molecule
• activated leukocyte cell adhesion moleculeFLJ38514
• ALCAM
• CD166 antigen
• CD166
• MEMDMGC71733

Background

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. The 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.

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ALCAM/CD166 Antibody (FAB6561P)(17)

Publications using FAB6561P

• YY Chun, KS Tan, L Yu, M Pang, MHM Wong, R Nakamoto, WZ Chua, A Huee-Ping, ZZR Lew, HH Ong, VT Chow, T Tran, D Yun Wang, LT Sham Influence of glycan structure on the colonization of Streptococcus pneumoniae on human respiratory epithelial cells Proceedings of the National Academy of Sciences of the United States of America, 2023-03-21;120(13):e2213584120. 3/21/2023 [PMID: 36943879]
• H Yoshida, L Koodie, K Jacobsen, K Hanzawa, Y Miyamoto, M Yamamoto B4GALNT1 induces angiogenesis, anchorage independence growth and motility, and promotes tumorigenesis in melanoma by induction of ganglioside GM2/GD2 Sci Rep, 2020-01-27;10(1):1199. 2020-01-27 [PMID: 31988291] (Flow Cytometry, Human)
• S Mount, P Kanda, S Parent, S Khan, C Michie, L Davila, V Chan, RA Davies, H Haddad, D Courtman, DJ Stewart, DR Davis Physiologic expansion of human heart-derived cells enhances therapeutic repair of injured myocardium Stem Cell Res Ther, 2019-11-04;10(1):316. 2019-11-04 [PMID: 31685023] (Flow Cytometry, Human)
• Anthony Essex, Javier Pineda, Grishma Acharya, Hong Xin, James Evans, Elizabeth Iorns et al. Replication Study: Wnt activity defines colon cancer stem cells and is regulated by the microenvironment eLife 2019-06-19 [PMID: 31215867]
• Z Wang, LY Yip, JHJ Lee, Z Wu, HY Chew, PKW Chong, CC Teo, HY Ang, KLE Peh, J Yuan, S Ma, LSK Choo, N Basri, X Jiang, Q Yu, AM Hillmer, WT Lim, TKH Lim, A Takano, EH Tan, DSW Tan, YS Ho, B Lim, WL Tam Methionine is a metabolic dependency of tumor-initiating cells Nat. Med., 2019-05-06;25(5):825-837. 2019-05-06 [PMID: 31061538] (Flow Cytometry, Human)
• Liu D, DU L, Chen D, Ye Z, Duan H, Tu T, Feng J, Yang Y, Chen Q, Yan X Reduced CD146 expression promotes tumorigenesis and cancer stemness in colorectal cancer through activating Wnt/beta-catenin signaling. Oncotarget, 2016-06-28;7(26):40704-40718. 2016-06-28 [PMID: 27302922] (Flow Cytometry, Human)
• James Evans, Anthony Essex, Hong Xin, Nurith Amitai, Lindsey Brinton, Erin Griner et al. Registered report: Wnt activity defines colon cancer stem cells and is regulated by the microenvironment eLife 2015-08-19 [PMID: 26287525]
• Fujiwara K, Ohuchida K, Sada M, Horioka K, Ulrich C, Shindo K, Ohtsuka T, Takahata S, Mizumoto K, Oda Y, Tanaka M CD166/ALCAM expression is characteristic of tumorigenicity and invasive and migratory activities of pancreatic cancer cells. PLoS ONE, 2014-09-15;9(9):e107247. 2014-09-15 [PMID: 25221999] (Flow Cytometry, Human)
• Rowehl , Rebecca, Burke , Stephani, Bialkowska , Agnieszk, Pettet , Donald W, Rowehl , Leahana, Li , Ellen, Antoniou , Eric, Zhang , Yuanhao, Bergamaschi , Roberto, Shroyer , Kenneth, Ojima , Iwao, Botchkina , Galina I Establishment of highly tumorigenic human colorectal cancer cell line (CR4) with properties of putative cancer stem cells. PLoS ONE, 2014-06-12;9(6):e99091. 2014-06-12 [PMID: 24921652] (Flow Cytometry, Human)
• Tachezy M, Zander H, Wolters-Eisfeld G, Muller J, Wicklein D, Gebauer F, Izbicki J, Bockhorn M Activated leukocyte cell adhesion molecule (CD166): an "inert" cancer stem cell marker for non-small cell lung cancer? Stem Cells, 2014-06-01;32(6):1429-36. 2014-06-01 [PMID: 24501004] (Flow Cytometry, Human)
• Mirkina I, Hadzijusufovic E, Krepler C, Mikula M, Mechtcheriakova D, Strommer S, Stella A, Jensen-Jarolim E, Holler C, Wacheck V, Pehamberger H, Valent P Phenotyping of human melanoma cells reveals a unique composition of receptor targets and a subpopulation co-expressing ErbB4, EPO-R and NGF-R. PLoS ONE, 2014-01-29;9(1):e84417. 2014-01-29 [PMID: 24489649] (Flow Cytometry, Human)
• Kalaszczynska Ilona, Ruminski Slawomir, Platek Anna E et al. Substantial differences between human and ovine mesenchymal stem cells in response to osteogenic media: how to explain and how to manage? BioResearch Open Access 1/1/2013 [PMID: 24083091]
• Albert R, Vereb Z, Csomos K, Moe M, Johnsen E, Olstad O, Nicolaissen B, Rajnavolgyi E, Fesus L, Berta A, Petrovski G Cultivation and characterization of cornea limbal epithelial stem cells on lens capsule in animal material-free medium. PLoS ONE, 2012-10-09;7(10):e47187. 2012-10-09 [PMID: 23056608] (Flow Cytometry, Human)
• Soh , Boon Sen, Zheng , Dahai, Li Yeo , Julie Su, Yang , Henry He, Ng , Shi Yan, Wong , Lan Hion, Zhang , Wencai, Li , Pin, Nichane , Massimo, Asmat , Atasha, Wong , Poo Sing, Wong , Peng Che, Su , Lin Lin, Mantalaris , Sakis A, Lu , Jia, Xian , Wa, McKeon , Frank, Chen , Jianzhu, Lim , Elaine H, Lim , Bing CD166(pos) subpopulation from differentiated human ES and iPS cells support repair of acute lung injury. Mol Ther, 2012-09-11;20(12):2335-46. 2012-09-11 [PMID: 22968480] (Cell Selection, Flow Cytometry, Human)
• Kondo S, Wakisaka N, Muramatsu M Epstein-Barr virus latent membrane protein 1 induces cancer stem/progenitor-like cells in nasopharyngeal epithelial cell lines. J. Virol., 2011-08-17;85(21):11255-64. 2011-08-17 [PMID: 21849440] (Flow Cytometry, Human)
• Giuliani M, Fleury M, Vernochet A, Ketroussi F, Clay D, Azzarone B, Lataillade JJ, Durrbach A Long-lasting inhibitory effects of fetal liver mesenchymal stem cells on T-lymphocyte proliferation. 2011-05-19;6(5):e19988. 2011-05-19 [PMID: 21625521] (Flow Cytometry, Human)
• Firth AL, Yao W, Ogawa A, Madani MM, Lin GY, Yuan JX Multipotent mesenchymal progenitor cells are present in endarterectomized tissues from patients with chronic thromboembolic pulmonary hypertension. Am. J. Physiol., Cell Physiol., 2010-02-24;298(5):C1217-25. 2010-02-24 [PMID: 20181931] (Flow Cytometry, ICC, Human)

Bioinformatics

• Gene Symbol: ALCAM
• Uniprot:
  • Q13740()