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AB261772

Human ATG16L1 knockout HeLa cell line

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ATG16L1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 22 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View 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

4 Images
Western blot - Human ATG16L1 knockout HeLa cell line (AB261772)
  • WB

Unknown

Western blot - Human ATG16L1 knockout HeLa cell line (AB261772)

Lanes 1-4 : Merged signal (red and green). Green - ab233796 observed at 68 and 72 kDa. Red - loading control ab181602 observed at 37 kDa.

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 ab256843) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab233796 and 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 (ab216772) and 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] (<a href='/en-us/products/primary-antibodies/atg16l1-antibody-5h9a11-ab233796'>ab233796</a>) 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

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution

Predicted band size: 68 kDa

Observed band size: 68 kDa,72 kDa

false

Western blot - Human ATG16L1 knockout HeLa cell line (AB261772)
  • WB

Supplier Data

Western blot - Human ATG16L1 knockout HeLa cell line (AB261772)

Lanes 1-4 : Merged signal (red and green). Green - ab187671 observed at 68 and 72 kDa. Red - loading control ab8245 observed at 37 kDa.

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 ab256843) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and 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 (ab216773) and 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 (<a href='/en-us/products/primary-antibodies/atg16l1-antibody-epr15638-n-terminal-ab187671'>ab187671</a>) 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

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 68 kDa

Observed band size: 68 kDa,72 kDa

false

Sanger Sequencing - Human ATG16L1 knockout HeLa cell line (AB261772)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG16L1 knockout HeLa cell line (AB261772)

Allele-2 : 22 bp deletion in exon 1.

Sanger Sequencing - Human ATG16L1 knockout HeLa cell line (AB261772)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG16L1 knockout HeLa cell line (AB261772)

Allele-1 : Insertion of the selection cassette in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 22 bp deletion in exon 1 and Insertion of the selection cassette in exon 1

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

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.

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What's included?

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Properties and storage information

Gene name
ATG16L1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information ATG16L1
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Product promise

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