Recombinant Anti-Axl antibody [EPR21107] (ab215205)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21107] to Axl
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Axl antibody [EPR21107]
See all Axl primary antibodies -
Description
Rabbit monoclonal [EPR21107] to Axl -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: DU 145, NCI-H1299, 4T1, C2C12, HeLa and HUVEC whole cell lysates; Human skeletal muscle lysate; Rat and mouse muscle lysates.
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21107 -
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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KO cell lines
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab215205 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 |
1/1000. Detects a band of approximately 80, 140 kDa (predicted molecular weight: 98 kDa).
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Notes |
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WB
1/1000. Detects a band of approximately 80, 140 kDa (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
- Entrez Gene: 26362 Mouse
- Entrez Gene: 308444 Rat
- Omim: 109135 Human
- SwissProt: P30530 Human
- SwissProt: Q00993 Mouse
- Unigene: 590970 Human
- Unigene: 4128 Mouse
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Alternative names
- Adhesion related kinase antibody
- AI323647 antibody
- Ark antibody
see all
Images
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All lanes : Anti-Axl antibody [EPR21107] (ab215205) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : AXL knockout A549 cell lysate
Lane 3 : A549 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 135 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Axl antibody [EPR21107] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab215205 was shown to bind specifically to Axl. A band was observed at 135 kDa in wild-type A549 cell lysates with no signal observed at this size in AXL knockout cell line ab273744 (knockout cell lysate ab273780). To generate this image, wild-type and AXL knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-Axl antibody [EPR21107] (ab215205) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate at 40 µg
Lane 2 : AXL knockout HeLa cell lysate at 40 µg
Lane 3 : Wild-type HeLa cell lysate at 20 µg
Lane 4 : AXL knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 135 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab215205 observed at 135 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab215205 was shown to react with Axl in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab215205 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Axl antibody [EPR21107] (ab215205) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : AXL knockout HeLa cell lysate
Lane 3 : NCI-H1299 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 80,140 kDa why is the actual band size different from the predicted?Western blot: Anti-Axl antibody [EPR21107] (ab215205) staining at 1:1000 dilution, shown in green; Anti-GAPDH antibody [6C5] - Loading Control (ab8245) shown in red.
ab215205 was shown to bind specifically to Axl. A band was observed at 140 kDa in wild-type HeLa cell lysates with no signal observed at this size in Axl CRISPR-Cas9 edited cell line ab265392 (CRISPR-Cas9 edited cell lysate ab257152). The band observed in the CRISPR-Cas9 edited lysate lane below 90 kDa is likely to represent a truncated form of Axl. This has not been investigated further and the functional properties of the gene product have not been determined.
Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/10000 dilution.
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All lanes : Anti-Axl antibody [EPR21107] (ab215205) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : AXL knockout HeLa cell lysate
Lane 3 : NCI-H1299 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 80,140 kDa why is the actual band size different from the predicted?Western blot: Anti-Axl antibody [EPR21107] (ab215205) staining at 1:1000 dilution, shown in green; Anti-GAPDH antibody [6C5] - Loading Control (ab8245) shown in red.
ab215205 was shown to bind specifically to Axl. A band was observed at 140 kDa in wild-type HeLa cell lysates with no signal observed at this size in Axl CRISPR-Cas9 edited cell line ab261810 (CRISPR-Cas9 edited cell lysate ab257151). The band observed in the CRISPR-Cas9 edited lysate lane below 90 kDa is likely to represent a truncated form of Axl. This has not been investigated further and the functional properties of the gene product have not been determined.
Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/10000 dilution.
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All lanes : Anti-Axl antibody [EPR21107] (ab215205) at 1/1000 dilution
Lane 1 : DU 145 (human prostate carcinoma cell line) whole cell lysate
Lane 2 : NCI-H1299 (human lung carcinoma cell line) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Predicted band size: 98 kDa
Observed band size: 140,80 kDa why is the actual band size different from the predicted?
Exposure time: 29 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Negative control: Jurkat
The 140 kDa band corresponds to the full-length Axl, while the 80 kDa band could be the cleaved form of Axl (PMID: 7822279 and 19541935).The apparent molecular mass is higher than predicted, likely due to glycosylation (PMID: 23629654).
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All lanes : Anti-Axl antibody [EPR21107] (ab215205) at 1/1000 dilution
Lane 1 : Human skeletal muscle lysate at 20 µg
Lane 2 : Rat muscle lysate at 10 µg
Lane 3 : Mouse muscle lysate at 10 µg
Lane 4 : 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate at 20 µg
Lane 5 : C2C12 (mouse myoblast cell line) whole cell lysate at 20 µg
Lane 6 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 7 : HUVEC (human umbilical vein endothelial cell line) whole cell lysate at 20 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Predicted band size: 98 kDa
Observed band size: 140,80 kDa why is the actual band size different from the predicted?Exposure time : Lane 1: 6 seconds; Lane 2: 3 seconds; Lane 3: 46 seconds; Lanes 4-5: 3 minutes; Lanes 6-7: 10 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The 140 kDa band corresponds to the full-length Axl, while the 80 kDa band could be the cleaved form of Axl (PMID: 7822279 and 19541935).
The apparent molecular mass is higher than predicted, likely due to glycosylation (PMID: 23629654).
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
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SDS download
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Datasheet download
Certificate of Compliance
References (6)
ab215205 has been referenced in 6 publications.
- Qian C et al. Opening KATP channels induces inflammatory tolerance and prevents chronic pain. Brain Behav Immun 107:76-86 (2023). PubMed: 36198341
- Xia B et al. Extracellular vesicles mediate antibody-resistant transmission of SARS-CoV-2. Cell Discov 9:2 (2023). PubMed: 36609376
- Scherschinski L et al. Regulation of the Receptor Tyrosine Kinase AXL in Response to Therapy and Its Role in Therapy Resistance in Glioblastoma. Int J Mol Sci 23:N/A (2022). PubMed: 35055167
- Zhang S et al. Bilirubin Improves Gap Junction to Alleviate Doxorubicin-Induced Cardiotoxicity by Regulating AMPK-Axl-SOCS3-Cx43 Axis. Front Pharmacol 13:828890 (2022). PubMed: 35548345
- Zhou G & Li H AXL Inhibits Proinflammatory Factors to Relieve Rheumatoid Arthritis Pain by Regulating the TLR4/NF-κB Signaling Pathway. Evid Based Complement Alternat Med 2022:7625739 (2022). PubMed: 35983008
- Fisher MJ et al. Cabozantinib for neurofibromatosis type 1-related plexiform neurofibromas: a phase 2 trial. Nat Med 27:165-173 (2021). PubMed: 33442015