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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD166 antibody [EPR2759(2)] (ab109215)

Reviews (3) Submit a question References (22)
Flow Cytometry - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)
  • Anti-CD166 antibody [EPR2759(2)] (ab109215)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2759(2)] to CD166
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free HRP PE

Overview

  • Product name

    Anti-CD166 antibody [EPR2759(2)]
    See all CD166 primary antibodies

  • Description

    Rabbit monoclonal [EPR2759(2)] to CD166

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: SH-SY5Y, HuT-78, HT1080, Daudi and HeLa whole cell lysate (ab150035). Mouse and rat brain tissue lysates; Wild-type HAP1 whole cell lysate. IHC-P: Human liver and prostatic adenocarcinoma tissues. ICC/IF: THP-1 cells. Flow Cyt: HuT-78 cells. IP: SH-SY5Y cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109215 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (1)
1/10000 - 1/20000. Detects a band of approximately 100-105 kDa (predicted molecular weight: 65 kDa).

For unpurified use at 1/1000 - 1/10000.
IP (1)
1/30.

For unpurified use at 1/10 - 1/100.
IHC-P (1)
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.
Flow Cyt
1/90.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/50 - 1/250.
Notes
WB
1/10000 - 1/20000. Detects a band of approximately 100-105 kDa (predicted molecular weight: 65 kDa).

For unpurified use at 1/1000 - 1/10000.
IP
1/30.

For unpurified use at 1/10 - 1/100.
IHC-P
1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/100 - 1/250.
Flow Cyt
1/90.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/50 - 1/250.

Target

  • Function

    Cell adhesion molecule that binds to CD6. Involved in neurite extension by neurons via heterophilic and homophilic interactions. May play a role in the binding of T- and B-cells to activated leukocytes, as well as in interactions between cells of the nervous system.

  • Tissue specificity

    Spleen, placenta, liver, and weakly in liver. Expressed by activated T-cells, B-cells, monocytes and thymic epithelial cells. Expressed by neurons in the brain. Restricted expression in tumor cell lines. Preferentially expressed in highly metastasizing melanoma cell lines.

  • Sequence similarities

    Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 2 Ig-like V-type (immunoglobulin-like) domains.

  • Domain

    The CD6 binding site is located in the N-terminal Ig-like domain.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Activated leukocyte cell adhesion molecule antibody
    • ALCAM antibody
    • ALCAM protein antibody
    • CD 166 antibody
    • CD166 antibody
    • CD166 antigen antibody
    • CD166_HUMAN antibody
    • FLJ3851 antibody
    • FLJ38514 antibody
    • MEMD antibody
    • MGC71733 antibody
    see all

Images

  • Flow Cytometry - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Flow Cytometry - Anti-CD166 antibody [EPR2759(2)] (ab109215)

    Flow Cytometry analysis of HuT-78 cells labelling CD166 with purified ab109215 at 1/90 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunocytochemistry/ Immunofluorescence - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Immunocytochemistry/ Immunofluorescence - Anti-CD166 antibody [EPR2759(2)] (ab109215)

    Immunocytochemistry/Immunofluorescence analysis of THP-1 (human monocytic leukemia cell line) cells labelling CD166 (green) with purified ab109215 at 1/250. Cells were fixed with 100% methanol. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as a nuclear counterstain.  

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with purified ab109215 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: CD166 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: SH-SY5Y whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109215 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab109215 was shown to specifically react with CD166 in wild-type HAP1 cells as signal was lost in CD166 knockout cells. Wild-type and CD166 knockout samples were subjected to SDS-PAGE. Ab109215 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Immunoprecipitation - Anti-CD166 antibody [EPR2759(2)] (ab109215)

    ab109215 (purified) at 1/30 immunoprecipitating CD166 in SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    All lanes : Anti-CD166 antibody [EPR2759(2)] (ab109215) at 1/10000 dilution (purified)

    Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
    Lane 2 : HuT-78 cell lysate
    Lane 3 : HT-1080 (human fibrosarcoma cell line) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 65 kDa
    Observed band size: 100-105 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    All lanes : Anti-CD166 antibody [EPR2759(2)] (ab109215) at 1/10000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 65 kDa
    Observed band size: 100-105 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD166 with unpurified ab109215 at 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Western blot - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    All lanes : Anti-CD166 antibody [EPR2759(2)] (ab109215) at 1/1000 dilution (unpurified)

    Lane 1 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
    Lane 2 : HuT-78 cell lysate
    Lane 3 : HT1080 (human fibrosarcoma cell line) cell lysate
    Lane 4 : Daudi (human Burkitt's lymphoma cell line) cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 65 kDa
    Observed band size: 100-105 kDa why is the actual band size different from the predicted?

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD166 antibody [EPR2759(2)] (ab109215)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic adenocarcinoma tissue labelling CD166 with unpurified ab109215 at 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Anti-CD166 antibody [EPR2759(2)] (ab109215)
    Anti-CD166 antibody [EPR2759(2)] (ab109215)

References (22)

Publishing research using ab109215? Please let us know so that we can cite the reference in this datasheet.

ab109215 has been referenced in 22 publications.

  • Pitts HA  et al. SPINK2 Protein Expression Is an Independent Adverse Prognostic Marker in AML and Is Potentially Implicated in the Regulation of Ferroptosis and Immune Response. Int J Mol Sci 24:N/A (2023). PubMed: 37298647
  • Kalantari E  et al. Significant co-expression of putative cancer stem cell markers, EpCAM and CD166, correlates with tumor stage and invasive behavior in colorectal cancer. World J Surg Oncol 20:15 (2022). PubMed: 35016698
  • Qian Q  et al. Intrahepatic activated leukocyte cell adhesion molecule induces CD6highCD4+ T cell infiltration in autoimmune hepatitis. Front Immunol 13:967944 (2022). PubMed: 36159854
  • Song J  et al. Cancer Stem Cell Formation Induced and Regulated by Extracellular ATP and Stanniocalcin-1 in Human Lung Cancer Cells and Tumors. Int J Mol Sci 23:N/A (2022). PubMed: 36499099
  • Grönroos P  et al. Directed Differentiation of Human Pluripotent Stem Cells towards Corneal Endothelial-Like Cells under Defined Conditions. Cells 10:N/A (2021). PubMed: 33562615
View all Publications for this product

Customer reviews and Q&As

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Immunoprecipitation abreview for Anti-CD166 antibody [EPR2759(2)]

 Excellent
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (melanoma 4L cell line)
Total protein in input
1000 µg
Immuno-precipitation step
Protein G
Specification
melanoma 4L cell line
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted May 12 2017

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