Human AXL knockout HeLa cell lysate
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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.
View Alternative Names
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
- WB
Unknown
Western blot - Human AXL knockout HeLa cell lysate (AB257151)
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)
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.
All lanes:
Western blot - Anti-Axl antibody [EPR21107] (<a href='/en-us/products/primary-antibodies/axl-antibody-epr21107-ab215205'>ab215205</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human AXL knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-axl-knockout-hela-cell-line-ab261810'>ab261810</a>)
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
Predicted band size: 98 kDa
Observed band size: 80140 kDa
false
- WB
Lab
Western blot - Human AXL knockout HeLa cell lysate (AB257151)
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 - 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).
All lanes:
Western blot - Anti-Axl antibody [EPR23892-15] (<a href='/en-us/products/primary-antibodies/axl-antibody-epr23892-15-ab259831'>ab259831</a>) 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
Secondary
All lanes:
Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution
Predicted band size: 98 kDa
Observed band size: 140 kDa,80 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human AXL knockout HeLa cell lysate (AB257151)
Allele-1 : 1 bp deletion in exon4
- Sanger seq
Unknown
Sanger Sequencing - Human AXL knockout HeLa cell lysate (AB257151)
Allele-2 : 4 bp deletion in exon4
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
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.
Pathways
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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