Human AXL knockout A549 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human AXL knockout A549 cell line (AB273744)
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.
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 A549 cell lysate at 20 µg
Lane 2:
AXL knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human AXL knockout A549 cell line (ab273744)
Lane 3:
A549 cell lysate at 20 µg
Lane 4:
Jurkat cell lysate at 20 µg
Predicted band size: 98 kDa
Observed band size: 135 kDa
false
- NGS
Lab
Next Generation Sequencing - Human AXL knockout A549 cell line (AB273744)
19 bp deletion after Gly38 of the WT protein
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Axl antibody [EPR19880] (AB219651)](https://content.abcam.com/products/images/axl-antibody-epr19880-ab219651--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img69874.jpg)
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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
Zygosity
Shipped at conditions
Appropriate short-term storage duration
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com