Recombinant Anti-Axl antibody [EPR23892-15] (ab259831)
![Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)](https://www.abcam.com/ps/products/259/ab259831/Images/ab259831-449771-anti-axlantibody-epr2389215-western-blot-epithelial.jpg "Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23892-15] to Axl
- Suitable for: ICC, Flow Cyt, WB
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-Axl antibody [EPR23892-15]
See all Axl primary antibodies -
Description
Rabbit monoclonal [EPR23892-15] to Axl -
Host species
Rabbit -
Tested applications
Suitable for: ICC, Flow Cyt, WBmore details
Unsuitable for: IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment within Human Axl aa 1-450. The exact sequence is proprietary.
Database link: P30530 -
Positive control
- WB: Wild-type HeLa whole cell lysate; NCI-H1299 whole cell lysate; DU145 whole cell lysate. ICC: NCI-H1299; Hela cells. Flow Cyt: Hela cells; NCI-H1299 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.
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
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23892-15 -
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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Related Products
Applications
Our Abpromise guarantee covers the use of ab259831 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 |
|---|---|---|
| ICC | 1/100. | |
| Flow Cyt | 1/500. | |
| WB | 1/1000. Predicted molecular weight: 98 kDa. |
Target
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Function
May function as a signal transducer between specific cell types of mesodermal origin. In case of filovirus infection, seems to function as a cell entry factor. -
Tissue specificity
Highly expressed in metastatic colon tumors. Expressed in primary colon tumors. Weakly expressed in normal colon tissue. -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. AXL/UFO subfamily.
Contains 2 fibronectin type-III domains.
Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 protein kinase domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 558 Human
- Omim: 109135 Human
- SwissProt: P30530 Human
- Unigene: 590970 Human
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Alternative names
- Adhesion related kinase antibody
- AI323647 antibody
- Ark antibody
see all
Images
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Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)All lanes : Anti-Axl antibody [EPR23892-15] (ab259831) at 1/1000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Axl knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate
Lane 4 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Predicted band size: 98 kDa
Observed band size: 140,80 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lanes 1-4: Merged signal (red and green). Green - ab259831 observed at 140, 80 kDa. Red - loading control ab8245 observed at 36 kDa.
ab259831 Anti-Axl antibody [EPR23892-15] was shown to react with Axl in Hela cells in Western blot. Loss of signal was observed when knockout cell line ab261810 (knockout cell lysate ab257151) was used. Wild-type and Axl knockout samples were subjected to SDS-PAGE.
ab259831 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
Negative control: Jurkat (PMID: 28423548).
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![Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)](https://www.abcam.com/ps/products/259/ab259831/Images/ab259831-1-anti-axlantibody-epr2389215-immunocytochemistry-ncih1299.jpg)
Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Trito nX-100 permeabilized NCI-H1299 cells labelling Axl with ab259831 at 1/500 (0.98 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong membranous and weak cytoplasmic staining in NCI-H1299 cells (PMID: 27443522). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Negative control: Jurkat (PMID: 28423548).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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![Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)](https://www.abcam.com/ps/products/259/ab259831/Images/ab259831-3-anti-axlantibody-epr2389215-flow-cytometry-parental-hela.jpg)
Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)Flow cytometric analysis of Parental HeLa (human cervix adenocarcinoma epithelial cell (Right panel)/ AXL knockout HeLa cells (Left panel) cells labelling Axl with ab259831 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Positive staining on HeLa cells (ab255928), while no staining on AXL knockout HeLa cells (ab261810).
Gated on viable cells.
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![Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)](https://www.abcam.com/ps/products/259/ab259831/Images/ab259831-449774-anti-axlantibody-epr2389215-western-blot-adenocarcinoma.jpg)
Western blot - Anti-Axl antibody [EPR23892-15] (ab259831)All lanes : Anti-Axl antibody [EPR23892-15] (ab259831) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : DU145 (human prostate carcinoma epithelial cell), whole cell lysate
Lane 3 : NCI-H1299 (human lung carcinoma epithelial cell), whole cell lysate
Lane 4 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 140,80 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: Jurkat (PMID: 28423548).
Exposure time: 37 seconds
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![Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)](https://www.abcam.com/ps/products/259/ab259831/Images/ab259831-2-anti-axlantibody-epr2389215-immunocytochemistry-axl-ko-hela.jpg)
Immunocytochemistry - Anti-Axl antibody [EPR23892-15] (ab259831)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Axl KO HeLa (ab261810) cells labelling Axl with ab259831 at 1/100 (4.9 µg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong membranous and weak cytoplasmic staining in parental HeLa cells (ab255928), while no staining in AXL knockout HeLa cells (ab261810). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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![Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)](https://www.abcam.com/ps/products/259/ab259831/Images/ab259831-4-anti-axlantibody-epr2389215-flow-cytometry-jurkat.jpg)
Flow Cytometry - Anti-Axl antibody [EPR23892-15] (ab259831)Flow cytometric analysis of Jurkat (Human T cell leukemia T lymphocyte, Left panel)/ NCI-H1299 (human lung carcinoma epithelial cell, Right panel) cells labelling Axl with ab259831 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Negative control: Jurkat (PMID: 28423548).
Gated on viable cells.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
Certificate of Compliance
References (0)
ab259831 has not yet been referenced specifically in any publications.
