Recombinant Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671)
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
Related conjugates and formulations
Overview
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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. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, Raji, Jurkat, Daudi, PC12 and NIH 3T3 cell lysates, Wild-type THP-1 cell lysate, Wild type HeLa cell lysate IHC-P: Human prostatic hyperplasia and Mouse colon tissues.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR15638 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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WB | (4) |
1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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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
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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
modificationsProteolytic 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
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Database links
- Entrez Gene: 55054 Human
- Entrez Gene: 77040 Mouse
- Entrez Gene: 363278 Rat
- Omim: 610767 Human
- SwissProt: Q676U5 Human
- SwissProt: Q8C0J2 Mouse
- Unigene: 529322 Human
- Unigene: 272972 Mouse
see all -
Form
There are 4 isoforms produced by alternative splicing. -
Alternative names
- A16L1_HUMAN antibody
- APG16 like 1 antibody
- APG16-like 1 antibody
see all
Images
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All lanes : Anti-ATG16L1 antibody [EPR15638] - N-terminal (ab187671) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : ATG16L1 knockout THP-1 cell lysate
Lane 3 : Wild type HeLa cell lysate
Lane 4 : ATG16L1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 68,70 kDa why is the actual band size different from the predicted?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] (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 ab277834 (knockout 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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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 kDaLanes 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.
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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.
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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.
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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: 68,72 kDa why is the actual band size different from the predicted?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.
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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: 68,72 kDa why is the actual band size different from the predicted?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.
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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 kDaLanes 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. -
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 -
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)
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (14)
ab187671 has been referenced in 14 publications.
- Zhang C et al. Autophagic sequestration of SQSTM1 disrupts the aggresome formation of ubiquitinated proteins during proteasome inhibition. Cell Death Dis 13:615 (2022). PubMed: 35840557
- Zhang L et al. Type I interferon signaling mediates Mycobacterium tuberculosis-induced macrophage death. J Exp Med 218:N/A (2021). PubMed: 33125053
- Chang HC et al. The BAX-binding protein MOAP1 associates with LC3 and promotes closure of the phagophore. Autophagy 17:3725-3739 (2021). PubMed: 33783314
- Li Y et al. The lncRNA HOTAIR regulates autophagy and affects lipopolysaccharide-induced acute lung injury through the miR-17-5p/ATG2/ATG7/ATG16 axis. J Cell Mol Med 25:8062-8073 (2021). PubMed: 34180119
- Xie S et al. miR-1307 promotes hepatocarcinogenesis by CALR-OSTC-endoplasmic reticulum protein folding pathway. iScience 24:103271 (2021). PubMed: 34761190