ATG16L1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided.
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Key facts
- Cell type
THP-1
- Species or organism
Human
- Tissue
Blood
- Form
Liquid
- Knockout validation
Western blot
Alternative names
A16L1_HUMAN, APG16-like 1, APG16L, APG16L beta, ATG16 autophagy related 16 like 1, ATG16 autophagy related 16-like 1 (S. cerevisiae), ATG16A, ATG16L, Atg16l1, Autophagy-related protein 16-1, FLJ00045, FLJ10035, FLJ10828, FLJ22677, IBD10, OTTHUMP00000164391, OTTHUMP00000164393, OTTHUMP00000165876, OTTHUMP00000165877, WD repeat domain 30, WDR30
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ATG16L1 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
ATG16L1
- 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
Recommended control: Human wild-type THP-1 cell line (Human wild-type THP-1 cell line ab281894). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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
ATG16L1 also known as Autophagy Related 16 Like 1 is a protein involved in the autophagy process. It functions as part of a complex that includes ATG12 and ATG5. The molecular weight of ATG16L1 is approximately 66 kDa. It is highly expressed in various tissues including the immune cells highlighting its involvement in essential cellular processes. The protein interacts with other autophagy-related proteins to facilitate the elongation and maturation of autophagosomes.
- Biological function summary
ATG16L1 plays a role in mediating autophagy a vital cellular degradation process. It forms a complex with ATG5 and ATG12 necessary for the elongation of the autophagosome membrane. Apart from its role in autophagy ATG16L1 contributes to the regulation of innate immunity by influencing the secretion of inflammatory cytokines. Its presence is essential for maintaining cellular homeostasis and proper immune responses.
- Pathways
ATG16L1 is important in pathways like autophagy and immunity. In the autophagy pathway it works alongside ATG5 and ATG12 to ensure the proper formation of autophagosomes which are structures that engulf and degrade unwanted cellular components. Furthermore in the immune response pathway it helps regulate inflammation by managing cytokine production and secretion showing interaction with proteins like NOD2.
- Associated diseases and disorders
ATG16L1 has connections with inflammatory bowel diseases particularly Crohn's disease and various cancers. Mutations in ATG16L1 can lead to impaired autophagy contributing to the development of Crohn's disease. In cancer dysregulation of autophagy involving ATG16L1 may affect tumor progression and response to therapy. The protein associates with NOD2 in Crohn's disease demonstrating that changes in their interaction can influence disease susceptibility and severity.
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2 product images
Western blot - Human ATG16L1 knockout THP-1 cell line (ab277834)
False colour image of Western blot: Anti-ATG16L1 antibody [EPR15638] - N-terminal staining at 1/1000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-ATG16L1 antibody [EPR15638] - N-terminal ab187671 was shown to bind specifically to ATG16L1. A band was observed at 68/70 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG16L1 knockout cell line ab277834 (knockout cell lysate Human ATG16L1 knockout THP-1 cell lysate ab278184). To generate this image wild-type and ATG16L1 knockout THP-1 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 °. 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 1/20000 dilution.
All lanes: Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (Anti-ATG16L1 antibody [EPR15638] - N-terminal ab187671) at 1/1000 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: ATG16L1 knockout THP-1 cell lysate at 20 µg
Lane 3: Wild type HeLa cell lysate at 20 µg
Lane 4: ATG16L1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 68 kDa, 70 kDa
Western blot - Human ATG16L1 knockout THP-1 cell line (ab277834)
False colour image of Western blot: Anti-ATG16L1 antibody [5H9A11] staining at 1/500 dilution shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/20000 dilution shown in red. In Western blot Anti-ATG16L1 antibody [5H9A11] ab233796 was shown to bind specifically to ATG16L1. A band was observed at 68/70 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG16L1 knockout cell line ab277834 (knockout cell lysate Human ATG16L1 knockout THP-1 cell lysate ab278184). To generate this image wild-type and ATG16L1 knockout THP-1 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °. 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
All lanes: Western blot - Anti-ATG16L1 antibody [5H9A11] (Anti-ATG16L1 antibody [5H9A11] ab233796) at 1/500 dilution
Lane 1: Wild-type THP-1 cell lysate at 20 µg
Lane 2: ATG16L1 knockout THP-1 cell lysate at 20 µg
Lane 3: Wild type HeLa cell lysate at 20 µg
Lane 4: ATG16L1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 68 kDa, 70 kDa
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