Recombinant Anti-ATG3 antibody [EPR4801] (ab108251)
![Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)](https://www.abcam.com/ps/products/108/ab108251/Images/ab108251-393585-anti-atg3-antibody-epr4801-western-blot-human-knockout-lysate.jpg "Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4801] to ATG3
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-ATG3 antibody [EPR4801]
See all ATG3 primary antibodies -
Description
Rabbit monoclonal [EPR4801] to ATG3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK293T, K562, HL-60, HeLa or Jurkat whole cell lysate (ab7899). IHC-P: Human muscle tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
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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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR4801 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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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 ab108251 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 | (2) |
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).
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| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Perform antigen retrieval before commencing with IHC staining protocol. |
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| Flow Cyt |
1/2000.
|
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| ICC/IF | (3) |
1/100 - 1/250.
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| Notes |
|---|
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WB
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa). |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Perform antigen retrieval before commencing with IHC staining protocol. |
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Flow Cyt
1/2000. |
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ICC/IF
1/100 - 1/250. |
Target
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Function
E2-like enzyme involved in autophagy and mitochondrial homeostasis. Catalyzes the conjugation of ATG8-like proteins (GABARAP, GABARAPL1, GABARAPL2 or MAP1LC3A) to phosphatidylethanolamine (PE). PE-conjugation to ATG8-like proteins is essential for autophagy. Preferred substrate is MAP1LC3A. Also acts as an autocatalytic E2-like enzyme, catalyzing the conjugation of ATG12 to itself, ATG12 conjugation to ATG3 playing a role in mitochondrial homeostasis but not in autophagy. ATG7 (E1-like enzyme) facilitates this reaction by forming an E1-E2 complex with ATG3. -
Tissue specificity
Widely expressed, with a highest expression in heart, skeletal muscle, kidney, liver and placenta. -
Sequence similarities
Belongs to the ATG3 family. -
Post-translational
modificationsConjugated to ATG12 at Lys-243. ATG12-conjugation plays a role in regulation of mitochondrial homeostasis and cell death, while it is not involved in PE-conjugation to ATG8-like proteins and autophagy. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 64422 Human
- Entrez Gene: 67841 Mouse
- Entrez Gene: 171415 Rat
- Omim: 609606 Human
- SwissProt: Q9NT62 Human
- SwissProt: Q9CPX6 Mouse
- SwissProt: Q6AZ50 Rat
- Unigene: 477126 Human
see all -
Alternative names
- 2610016C12Rik antibody
- Apg 3 antibody
- APG3 antibody
see all
Images
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Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)All lanes : Anti-ATG3 antibody [EPR4801] (ab108251) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ATG3 knockout HEK293T cell lysate
Lane 3 : K-562 cell lysate
Lane 4 : HeLa 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: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab108251 observed at 40 kDa. Red - loading control ab7291 observed at 50 kDa.
ab108251 Anti-ATG3 antibody [EPR4801] was shown to specifically react with ATG3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266707 (knockout cell lysate ab257363) was used. Wild-type and ATG3 knockout samples were subjected to SDS-PAGE. ab108251 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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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![Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody [EPR4801] (ab108251)](https://www.abcam.com/ps/products/108/ab108251/Images/ab108251-120760-anti-atg3-antibody-epr4801-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody [EPR4801] (ab108251)Immunofluorescent staining of HeLa cells using ab108251 at 1/100. -
![Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)](https://www.abcam.com/ps/products/108/ab108251/Images/ab108251-120762-anti-atg3-antibody-epr4801-western-blot.jpg)
Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)All lanes : Anti-ATG3 antibody [EPR4801] (ab108251) at 1/10000 dilution
Lane 1 : K562 cell lysate
Lane 2 : HL-60 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted? -
![Flow Cytometry - Anti-ATG3 antibody [EPR4801] (ab108251)](https://www.abcam.com/ps/products/108/ab108251/Images/ab108251-280406-anti-atg3-antibody-epr4801-flow-cytometry.jpg)
Flow Cytometry - Anti-ATG3 antibody [EPR4801] (ab108251)Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATG3 (red) with ab108251 at a 1/2000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG3 antibody [EPR4801] (ab108251)](https://www.abcam.com/ps/products/108/ab108251/Images/ab108251-120761-anti-atg3-antibody-epr4801-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG3 antibody [EPR4801] (ab108251)ab108251, at a 1/100 dilution, staining Human ATG3 in muscle, using Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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![Anti-ATG3 antibody [EPR4801] (ab108251)](https://www.abcam.com/ps/products/108/ab108251/Images/ab108251-7-benefits-of-recombinant-antibodies.png)
Anti-ATG3 antibody [EPR4801] (ab108251)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (12)
ab108251 has been referenced in 12 publications.
- Li G et al. Long non-coding RNA growth arrest-specific 5 (GAS5) acts as a tumor suppressor by promoting autophagy in breast cancer. Mol Med Rep 22:2460-2468 (2020). PubMed: 32705220
- Qiu S et al. The blockade of lipophagy pathway is necessary for docosahexaenoic acid to regulate lipid droplet turnover in hepatic stellate cells. Biomed Pharmacother 109:1841-1850 (2019). PubMed: 30551439
- Agrotis A et al. Redundancy of human ATG4 protease isoforms in autophagy and LC3/GABARAP processing revealed in cells. Autophagy 15:976-997 (2019). PubMed: 30661429
- Stavoe AK et al. Expression of WIPI2B counteracts age-related decline in autophagosome biogenesis in neurons. Elife 8:N/A (2019). PubMed: 31309927
- Agrotis A et al. Human ATG4 autophagy proteases counteract attachment of ubiquitin-like LC3/GABARAP proteins to other cellular proteins. J Biol Chem 294:12610-12621 (2019). PubMed: 31315929