AXL KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon4 and 4 bp deletion in exon4.
AI323647, AXL oncogene, AXL receptor tyrosine kinase, AXL transforming gene, AXL transforming sequence/gene, Adhesion related kinase, Ark, EC 2.7.10.1, JTK11, Oncogene AXL, Tyro7, Tyrosine-protein kinase receptor UFO, UFO_HUMAN
AXL KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon4 and 4 bp deletion in exon4.
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Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
The Axl protein also known as AXL receptor tyrosine kinase plays a significant role in cell signaling. It has a molecular weight of approximately 98 kDa. Axl is a transmembrane receptor that is expressed in a variety of tissues including the immune system reproductive organs and the central nervous system. It is mainly recognized for transmitting signals from the extracellular matrix into the cytoplasm by binding with its ligand Gas6.
Axl is important in mediating cell survival proliferation and migration. It functions as part of the TAM family of receptors which also includes Tyro3 and Mer. Axl often forms complexes with these receptors to facilitate efficient signaling. This interaction triggers phosphorylation events that activate downstream signaling proteins reinforcing its influence in cellular functions.
Axl is deeply involved in the PI3K-Akt and MAPK signaling pathways which are central to cellular growth and survival. In these pathways Axl interacts closely with proteins like PI3K and Akt to regulate processes such as apoptosis and metabolism. Its interaction with these pathways places Axl as an important modulator of intracellular signaling cascades.
Axl has been implicated in cancer progression and immune response dysregulation. In many cancer types Axl overexpression links to tumor invasion metastasis and resistance to therapy. Moreover Axl’s role in autoimmune diseases becomes evident through its association with the Gas6 protein. Exploring these connections furthers understanding of how Axl's aberrant activity influences health and disease.
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Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lane 1: Wild-type HeLa cell lysate (20 μg)
Lane 2: AXL knockout HeLa cell lysate (20 μg)
Lane 3: NCI-H1299 cell lysate (20 μg)
Lane 4: Jurkat cell lysate (20 μg)
Lanes 1-4: Merged signal (red and green). Green - Anti-Axl antibody [EPR23892-15] ab259831 observed at 140, 80 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Axl antibody [EPR23892-15] 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 Human AXL knockout HeLa cell line ab261810 (knockout cell lysate ab257151) was used. Wild-type and Axl knockout samples were subjected to SDS-PAGE.
Anti-Axl antibody [EPR23892-15] ab259831 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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).
All lanes: Western blot - Anti-Axl antibody [EPR23892-15] (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 at 40 µg
Lane 2: Axl knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 40 µg
Lane 3: NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 40 µg
Lane 4: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 40 µg
All lanes: Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution
Predicted band size: 98 kDa
Observed band size: 140 kDa, 80 kDa
Lane 2: AXL knockout HeLa cell lysate (20 μg)
Lane 3: NCI-H1299 cell lysate (20 μg)
Lane 4: Jurkat cell lysate (20 μg)
Western blot: Anti-Axl antibody [EPR21107] (Anti-Axl antibody [EPR21107] ab215205) staining at 1:1000 dilution, shown in green; Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) shown in red.
Anti-Axl antibody [EPR21107] 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 Human AXL knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution.
All lanes: Western blot - Anti-Axl antibody [EPR21107] (Anti-Axl antibody [EPR21107] ab215205) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: Western blot - Human AXL knockout HeLa cell line (Human AXL knockout HeLa cell line ab261810)
Lane 2: Western blot - Human AXL knockout HeLa cell lysate (ab257151)
Lane 2: AXL knockout HeLa cell lysate at 20 µg
Lane 3: NCI-H1299 cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 98 kDa
Observed band size: 80140 kDa
Allele-2: 4 bp deletion in exon4
Allele-1: 1 bp deletion in exon4
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