ATG5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Western blot
Alternative names
APG 5, APG 5L, APG5 autophagy 5 like, APG5-like, ASP, ATG5 autophagy related 5 homolog, ATG5_HUMAN, Apoptosis-specific protein, Autophagy protein 5, Autophagy related 5, Homolog of S Cerevisiae autophagy 5, OTTHUMP00000040507, hAPG5
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ATG5 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- THP-1
- Species or organism
- Human
- Tissue
- Blood
- Form
- Liquid
- Knockout validation
- Western blot
- Disease
- Acute Monocytic Leukemia
- Concentration
Properties
- Gene name
- ATG5
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Suspension
- Gender
- Male
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
- Culture medium
- RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
APG5L commonly known as ATG5 or ATG5 protein plays a significant role in autophagy. This molecular component has a molecular weight of approximately 32 kDa. ATG5 is expressed in multiple tissue types with a higher expression profile in immune-related cells. The protein is essential for the formation of the autophagosome as it conjugates with ATG12 to form a complex that is critical for autophagosome membrane elongation.
- Biological function summary
ATG5 plays an important part in the cellular process of autophagy affecting the degradation and recycling of cellular components. It is a part of a complex with ATG12 and ATG16L1 which facilitates the membrane building of the autophagosome. This protein is important for maintaining cellular homeostasis allowing the removal of damaged proteins and organelles and is involved in responses to stress and starvation.
- Pathways
ATG5 is an important component in the autophagy pathway and is involved in cellular maintenance processes. This pathway is vital for the regulation of cell growth and metabolism. ATG5 works closely with other autophagy-related proteins such as LC3. The ATG5-ATG12 complex helps in the lipidation of LC3 which is critical for autophagosome maturation connecting its function to the mTOR signaling pathway that regulates many cellular processes.
- Associated diseases and disorders
Disruption in ATG5 function is linked to neurodegenerative diseases and cancer. For instance dysregulation of autophagy due to ATG5 dysfunction may result in the progression of neurodegenerative disorders like Alzheimer's disease. Additionally in certain cancers altered autophagy can influence the survival of cancer cells. The interplay between ATG5 and other proteins like p53 and Bcl-2 is under investigation to understand its role in these diseases.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
2 product images
Western blot - Human ATG5 knockout THP-1 cell line (ab277835)
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (Anti-APG5L/ATG5 antibody [EPR4797] ab109490) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: ATG5 knockout THP-1 cell lysate at 20 µg
Lane 3: U-87 MG cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
SecondaryLanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 50 kDa
Western blot - Human ATG5 knockout THP-1 cell line (ab277835)
All lanes: Western blot - Anti-APG5L/ATG5 antibody [EPR1755(2)] (Anti-APG5L/ATG5 antibody [EPR1755(2)] ab108327) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: ATG5 knockout THP-1 cell lysate at 20 µg
Lane 3: U-87 MG cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
SecondaryLanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 50 kDa
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