Validated using a knockout cell line
Recombinant
RabMAb

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

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15638] to ATG16L1 - N-terminal
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ATG16L1 antibody [EPR15638] - N-terminal
    See all ATG16L1 primary antibodies

  • Description

    Rabbit monoclonal [EPR15638] to ATG16L1 - N-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human ATG16L1 aa 1-150 (N terminal). The exact sequence is proprietary.
    Database link: Q676U5

  • Positive control

    • WB: HeLa, Raji, Jurkat, Daudi, PC12 and NIH 3T3 cell lysates. IHC-P: Human prostatic hyperplasia and Mouse colon tissues.

  • General notes

     

     

    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 production
    For more information see here.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab187671 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Plays an essential role in autophagy: interacts with ATG12-ATG5 to mediate the conjugation of phosphatidylethanolamine (PE) to LC3 (MAP1LC3A, MAP1LC3B or MAP1LC3C), to produce a membrane-bound activated form of LC3 named LC3-II. Thereby, controls the elongation of the nascent autophagosomal membrane.

  • Involvement in disease

    Inflammatory bowel disease 10

  • Sequence similarities

    Belongs to the WD repeat ATG16 family.
    Contains 7 WD repeats.

  • Post-translational
    modifications

    Proteolytic cleavage by activated CASP3 leads to degradation and may regulate autophagy upon cellular stress and apoptotic stimuli.

  • Cellular localization

    Cytoplasm. Preautophagosomal structure membrane. Recruited to omegasomes membranes by WIPI2. Omegasomes are endoplasmic reticulum connected strutures at the origin of preautophagosomal structures. Localized to preautophagosomal structure (PAS) where it is involved in the membrane targeting of ATG5. Localizes also to discrete punctae along the ciliary axoneme.
  • Information by UniProt

  • Database links

    see all

  • Form

    There are 4 isoforms produced by alternative splicing.

  • Alternative names

    • A16L1_HUMAN antibody
    • APG16 like 1 antibody
    • APG16-like 1 antibody
    • APG16L antibody
    • APG16L beta antibody
    • ATG16 autophagy related 16 like 1 antibody
    • ATG16 autophagy related 16-like 1 (S. cerevisiae) antibody
    • ATG16A antibody
    • ATG16L antibody
    • Atg16l1 antibody
    • Autophagy related protein 16 1 antibody
    • Autophagy-related protein 16-1 antibody
    • FLJ00045 antibody
    • FLJ10035 antibody
    • FLJ10828 antibody
    • FLJ22677 antibody
    • IBD10 antibody
    • OTTHUMP00000164391 antibody
    • OTTHUMP00000164393 antibody
    • OTTHUMP00000165876 antibody
    • OTTHUMP00000165877 antibody
    • WD repeat domain 30 antibody
    • WDR30 antibody
    see all

Images

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

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa
    Observed band size: 68 kDa



    Lanes 1- 4: Merged signal (red and green). Green - ab187671 observed at 68 kDa. Red - 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 ab265263 (knockout 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 (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 (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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)
    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)
    Western blot - Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)
    All lanes : Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 68 kDa
    Observed band size: 75 kDa
    why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab187671 observed at 75 kDa. Red - 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 ab265263 (knockout cell lysate ab256842) was used. Wild-type and ATG16L1 knockout samples were subjected to SDS-PAGE. ab187671 and 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 (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.

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

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa



    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 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 (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 (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.

     

     

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

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG16L1 knockout HeLa cell lysate
    Lane 3 : Jurkat cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 68 kDa



    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°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 (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.

     

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

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : ATG16L1 knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 68 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab187671 observed at 68 and 70 kDa. Red - 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

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

    Lane 1 : HeLa cell lysate
    Lane 2 : Raji cell lysate
    Lane 3 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Lane 1 : PC12 cell lysate
    Lane 2 : NIH 3T3 cell lysate

    Lysates/proteins at 10 µg per lane.

    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)
    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.

References (6)

Publishing research using ab187671? Please let us know so that we can cite the reference in this datasheet.

ab187671 has been referenced in 6 publications.

  • Stavoe AK  et al. Expression of WIPI2B counteracts age-related decline in autophagosome biogenesis in neurons. Elife 8:N/A (2019). PubMed: 31309927
  • Tian W  et al. An antibody for analysis of autophagy induction. Nat Methods N/A:N/A (2019). PubMed: 31768061
  • Xu Y  et al. A Bacterial Effector Reveals the V-ATPase-ATG16L1 Axis that Initiates Xenophagy. Cell 178:552-566.e20 (2019). PubMed: 31327526
  • Lin X  et al. Liver-specific deletion of Eva1a/Tmem166 aggravates acute liver injury by impairing autophagy. Cell Death Dis 9:768 (2018). PubMed: 29991758
  • Zhang HN  et al. Berberine promotes nerve regeneration through IGFR-mediated JNK-AKT signal pathway. Mol Med Rep 18:5030-5036 (2018). PubMed: 30272344
  • Fan X  et al. Foot-and-mouth disease virus infection suppresses autophagy and NF-?B antiviral responses via degradation of ATG5-ATG12 by 3C(pro). Cell Death Dis 8:e2561 (2017). PubMed: 28102839

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Western blot abreview for Anti-ATG16L1 antibody [EPR15638] - N-terminal

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
African green monkey Cell lysate - whole cell (Kidney)
Gel Running Conditions
Reduced Denaturing (4-12% gel)
Loading amount
20 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Oct 06 2017

Western blot abreview for Anti-ATG16L1 antibody [EPR15638] - N-terminal

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Brain)
Gel Running Conditions
Reduced Denaturing (4-12% gel)
Loading amount
20 µg
Specification
Brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Oct 06 2017

Western blot abreview for Anti-ATG16L1 antibody [EPR15638] - N-terminal

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Kidney)
Gel Running Conditions
Reduced Denaturing (4-12% gel)
Loading amount
20 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Oct 06 2017

Western blot abreview for Anti-ATG16L1 antibody [EPR15638] - N-terminal

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK)
Gel Running Conditions
Reduced Denaturing
Loading amount
1 µg
Specification
HEK
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More

Dr. Ryan Russell

Verified customer

Submitted Sep 02 2016

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