Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2759(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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 |
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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. |
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ICC/IF |
1/50 - 1/250.
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Notes |
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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
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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
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Database links
- Entrez Gene: 214 Human
- Entrez Gene: 11658 Mouse
- Entrez Gene: 79559 Rat
- Omim: 601662 Human
- SwissProt: Q13740 Human
- SwissProt: Q61490 Mouse
- SwissProt: O35112 Rat
- Unigene: 591293 Human
see all -
Alternative names
- Activated leukocyte cell adhesion molecule antibody
- ALCAM antibody
- ALCAM protein antibody
see all
Images
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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) 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (22)
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