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Rabbit Recombinant Monoclonal ATG16L1 antibody. N-terminal. Suitable for WB, IHC-P and reacts with Rat, Human, Mouse samples. Cited in 18 publications.


Images

Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (AB187671), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - N-terminal (AB187671), expandable thumbnail
  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (AB187671), expandable thumbnail
  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (AB187671), expandable thumbnail
  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (AB187671), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-P
Human
Tested
Tested
Mouse
Expected
Tested
Rat
Tested
Expected

Tested
Tested

Species

Rat

Dilution info

1/1000 - 1/10000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Human

Dilution info

1/1000 - 1/10000

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/50 - 1/100

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Species

Human

Dilution info

1/50 - 1/100

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

5 products for Alternative Product

Target data

Function

Plays an essential role in both canonical and non-canonical autophagy: interacts with ATG12-ATG5 to mediate the lipidation to ATG8 family proteins (MAP1LC3A, MAP1LC3B, MAP1LC3C, GABARAPL1, GABARAPL2 and GABARAP) (PubMed:23376921, PubMed:23392225, PubMed:24553140, PubMed:24954904, PubMed:27273576, PubMed:29317426, PubMed:30778222, PubMed:33909989). Acts as a molecular hub, coordinating autophagy pathways via distinct domains that support either canonical or non-canonical signaling (PubMed:29317426, PubMed:30778222). During canonical autophagy, interacts with ATG12-ATG5 to mediate the conjugation of phosphatidylethanolamine (PE) to ATG8 proteins, to produce a membrane-bound activated form of ATG8 (PubMed:23376921, PubMed:23392225, PubMed:24553140, PubMed:24954904, PubMed:27273576). Thereby, controls the elongation of the nascent autophagosomal membrane (PubMed:23376921, PubMed:23392225, PubMed:24553140, PubMed:24954904, PubMed:27273576). As part of the ATG8 conjugation system with ATG5 and ATG12, required for recruitment of LRRK2 to stressed lysosomes and induction of LRRK2 kinase activity in response to lysosomal stress (By similarity). Also involved in non-canonical autophagy, a parallel pathway involving conjugation of ATG8 proteins to single membranes at endolysosomal compartments, probably by catalyzing conjugation of phosphatidylserine (PS) to ATG8 (PubMed:33909989). Non-canonical autophagy plays a key role in epithelial cells to limit lethal infection by influenza A (IAV) virus (By similarity). Regulates mitochondrial antiviral signaling (MAVS)-dependent type I interferon (IFN-I) production (PubMed:22749352, PubMed:25645662). Negatively regulates NOD1- and NOD2-driven inflammatory cytokine response (PubMed:24238340). Instead, promotes an autophagy-dependent antibacterial pathway together with NOD1 or NOD2 (PubMed:20637199). Plays a role in regulating morphology and function of Paneth cell (PubMed:18849966).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal ATG16L1 antibody. N-terminal. Suitable for WB, IHC-P and reacts with Rat, Human, Mouse samples. Cited in 18 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR15638

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

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.

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:
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12 product images

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    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, 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 Human ATG16L1 knockout THP-1 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 °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 (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 (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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Immunohistochemical analysis of paraffin-embedded Human prostatic hyperplasia tissue labeling ATG16L1 with ab187671 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Lanes 1- 4: Merged signal (red and green). Green - ab187671 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab187671 was shown to react with ATG16L1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human ATG16L1 knockout HeLa cell line ab265263 (knockout cell lysate Human ATG16L1 knockout HeLa cell lysate ab256842) was used. Wild-type HeLa and ATG16L1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 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 (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 3: Jurkat cell lysate at 20 µg

    Lane 4: Daudi cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 68 kDa

    Observed band size: 68 kDa

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 68 and 72 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human ATG16L1 knockout HeLa 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. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C 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 (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 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 (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

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 75 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

    ab187671 Anti-ATG16L1 antibody [EPR15638] was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human ATG16L1 knockout HeLa cell line ab265263 (knockout cell lysate Human ATG16L1 knockout HeLa cell lysate ab256842) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours 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 (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 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution

    Predicted band size: 68 kDa

    Observed band size: 75 kDa

    This data was developed using the same antibody clone in a different buffer formulation (ab187671).

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 75 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

    ab187671 Anti-ATG16L1 antibody [EPR15638] was shown to specifically react with ATG16L1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human ATG16L1 knockout HeLa cell line ab265263 (knockout cell lysate Human ATG16L1 knockout HeLa cell lysate ab256842) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours 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.

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 68 and 72 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human ATG16L1 knockout HeLa cell line ab261773 (knockout cell lysate ab256844) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C 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 (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 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 (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

    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 68 and 72 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human ATG16L1 knockout HeLa cell line ab261773 (knockout cell lysate ab256844) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C 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.

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Lanes 1 - 4: Merged signal (red and green). Green - ab187671 observed at 68 and 70 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
    ab187671 was shown to recognize ATG16L1 when ATG16L1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/2000 and 10000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging.

    All lanes: Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/2000 dilution

    Lane 1: Wild-type HAP1 cell lysate at 20 µg

    Lane 2: ATG16L1 knockout HAP1 cell lysate at 20 µg

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: Jurkat cell lysate at 20 µg

    Predicted band size: 68 kDa

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    All lanes: Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/5000 dilution

    Lane 1: HeLa cell lysate at 20 µg

    Lane 2: Raji cell lysate at 20 µg

    Lane 3: Jurkat cell lysate at 20 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 68 kDa

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    All lanes: Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/1000 dilution

    Lane 1: PC12 cell lysate at 10 µg

    Lane 2: NIH 3T3 cell lysate at 10 µg

    Secondary

    All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 68 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling ATG16L1 with ab187671 at 1/100 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate.

    In Western blot, Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) staining at 1/1000 dilution.

    Anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.Anti-His antibody (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    Exposure time: 3 minutes.

    All lanes: Western blot - Anti-ATG16L1 (phospho S278) antibody [EPR19016] - Rat IgG1 Chimeric (Anti-ATG16L1 (phospho S278) antibody [EPR19016] - Rat IgG1 (Chimeric) ab309495) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) transfected with an empty vector (vector control) containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

    Lane 2: 293T cells transfected with a human ATG16L1 expression vector containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

    Lane 3: 293T cells transfected with a human ATG16L1 (mutation S278A) expression vector containing a myc-His-tag whole cell lysate (untreated membrane) at 20 µg

    Lane 4: 293T (human embryonic kidney epithelial cell) transfected with an empty vector (vector control) containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

    Lane 5: 293T cells transfected with a human ATG16L1 expression vector containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

    Lane 6: 293T cells transfected with a human ATG16L1 (mutation S278A) expression vector containing a myc-His-tag whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rat IgG H&L (HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/5000 dilution

    Observed band size: 68 kDa

  • Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671), expandable thumbnail

    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate.

    In Western blot, Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) staining at 1/1000 dilution.

    anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

    Exposure time: 3 minutes.

    All lanes: Western blot - Anti-ATG16L1 (phospho S278) antibody [EPR19016] - Rat IgG1 Chimeric (Anti-ATG16L1 (phospho S278) antibody [EPR19016] - Rat IgG1 (Chimeric) ab309495) at 1/1000 dilution

    Lane 1: HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

    Lane 2: HCT116 amino acid and serum starved in EBSS(HBSS) for 2h whole cell lysate (untreated membrane) at 20 µg

    Lane 3: HCT116 whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

    Lane 4: HCT116 amino acid and serum starved in EBSS(HBSS) for 2h whole cell lysate (Alkaline phosphatase treated membrane) at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rat IgG H&L (HRP) (Goat Anti-Rat IgG H&L (HRP) ab205720) at 1/1000 dilution

    Observed band size: 68 kDa

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