Recombinant Anti-CD6/T12 antibody [EPR4057] (ab109217)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4057] to CD6/T12
- Suitable for: WB, IP, IHC-P, Flow Cyt (Intra), ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD6/T12 antibody [EPR4057]
See all CD6/T12 primary antibodies -
Description
Rabbit monoclonal [EPR4057] to CD6/T12 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-P, Flow Cyt (Intra), ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, HuT-78, and Human peripheral blood mononuclear cells whole cell lysates. Human tonsil tissue lysate; IP: Human peripheral blood mononuclear cells whole cell lysate; ICC/IF: Human peripheral blood mononuclear cells; IHC-P: Human tonsil and Human lymphocytes infiltrating colon tissues; Flow Cyt (intra): HuT-78 cells.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4057 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109217 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 |
1/5000. Detects a band of approximately 90-130 kDa (predicted molecular weight: 72 kDa).
For unpurified use at 1/1000 - 1/10000. |
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IP |
1/20.
For unpurified use at 1/10 - 1/100. |
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
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Flow Cyt (Intra) |
1/20.
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ICC/IF |
1/50.
For unpurified use at 1/100 - 1/250. |
Notes |
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WB
1/5000. Detects a band of approximately 90-130 kDa (predicted molecular weight: 72 kDa). For unpurified use at 1/1000 - 1/10000. |
IP
1/20. For unpurified use at 1/10 - 1/100. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
Flow Cyt (Intra)
1/20. |
ICC/IF
1/50. For unpurified use at 1/100 - 1/250. |
Target
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Function
Involved in cell adhesion. Binds to CD166. -
Tissue specificity
Expressed by thymocytes, mature T-cells, a subset of B-cells known as B-1 cells, and by some cells in the brain. -
Sequence similarities
Contains 3 SRCR domains. -
Post-translational
modificationsAfter T-cell activation, becomes hyperphosphorylated on Ser and Thr residues and phosphorylated on Tyr residues.
Contains intrachain disulfide bond(s). -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 923 Human
- Omim: 186720 Human
- SwissProt: P30203 Human
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Alternative names
- CD_antigen=CD6 antibody
- CD6 antibody
- CD6 antigen Tp120 antibody
see all
Images
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All lanes : Anti-CD6/T12 antibody [EPR4057] (ab109217) at 1/1000 dilution (unpurified)
Lane 1 : Jurkat lysates
Lane 2 : Human tonsil lysates
Lane 3 : PBMC lysates
Lane 4 : HuT-78 lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 72 kDa
Observed band size: 90-130 kDa why is the actual band size different from the predicted? -
Immunocytochemistry/Immunofluorescence analysis of Human PBMC (Human primary peripheral blood mononuclear cell) cells labeling CD6/T12 with Purified ab109217 at 1:50 dilution (2.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) cells, labeling CD6/T12 with Purified ab109217 at 1/20 dilution (10µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Purified ab109217 at 1:20 dilution (0.6μg) immunoprecipitating CD6/T12 in HuT-78 whole cell lysate.
Lane 1 (input): HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate 10μg
Lane 2 (+): ab109217 + HuT-78 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109217 in HuT-78 whole cell lysate.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 105-130 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD6/T12 with purified ab109217 at 1/1000 dilution (0.115 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Anti-CD6/T12 antibody [EPR4057] (ab109217) at 1/5000 dilution (Purified) + HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 72 kDa
Observed band size: 105-130 kDa why is the actual band size different from the predicted?The molecular weight of CD6 ranges from 105 to 130 kDa depending on its degree of phosphorylation (PMID: 17601777).
Blocking/Diluting buffer: 5% NFDM/TBST
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Unpurified ab109217, at a 1/100 dilution, staining CD6/T12 in paraffin embedded Human tonsil tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab109217, at a 1/100 dilution, staining CD6/T12 in paraffin embedded Human lymphocytes infiltrating colon tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab109217 showing negative staining in Skeletal muscle tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab109217 showing negative staining in Normal brain tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab109217 showing negative staining in Normal kidney tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Unpurified ab109217 showing negative staining in Normal uterus tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab109217 has been referenced in 1 publication.
- 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