ATG16L1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 22 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
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
ATG16L1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 22 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
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.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
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.
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.
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.
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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Anti-ATG16L1 antibody [5H9A11] ab233796 Anti-ATG16L1 antibody [5H9A11] was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261772 (knockout cell lysate Human ATG16L1 knockout HeLa cell lysate ab256843) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. Anti-ATG16L1 antibody [5H9A11] ab233796 and Anti-GAPDH antibody[EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-ATG16L1 antibody [5H9A11] (Anti-ATG16L1 antibody [5H9A11] ab233796) at 1/500 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ATG16L1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human ATG16L1 knockout HeLa cell line (ab261772)
Lane 3: Jurkat cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 68 kDa, 72 kDa
Anti-ATG16L1 antibody [EPR15638] - N-terminal ab187671 Anti-ATG16L1 antibody [EPR15638] - N-terminal was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261772 (knockout cell lysate Human ATG16L1 knockout HeLa cell lysate ab256843) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. Anti-ATG16L1 antibody [EPR15638] - N-terminal ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° 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 20000 dilution for 1 hour at room temperature before imaging.
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 HeLa cell lysate at 20 µg
Lane 2: ATG16L1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human ATG16L1 knockout HeLa cell line (ab261772)
Lane 3: Jurkat cell lysate at 20 µg
Lane 4: Daudi cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 68 kDa, 72 kDa
Allele-2: 22 bp deletion in exon 1.
Allele-1: Insertion of the selection cassette in exon 1.
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