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Rabbit Recombinant Monoclonal ATG12 antibody. Suitable for IHC-P, WB and reacts with Human, Mouse, Rat samples. Cited in 8 publications.

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Images

Western blot - Anti-ATG12 antibody [EPR4800] (AB109491), expandable thumbnail
  • Western blot - Anti-ATG12 antibody [EPR4800] (AB109491), expandable thumbnail
  • Western blot - Anti-ATG12 antibody [EPR4800] (AB109491), expandable thumbnail
  • Western blot - Anti-ATG12 antibody [EPR4800] (AB109491), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR4800] (AB109491), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.05% Sodium azide
Constituents: 50% Tissue culture supernatant, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 0.1% 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
IHC-PICC/IFIPFlow CytWB
Human
Tested
Not recommended
Not recommended
Not recommended
Tested
Mouse
Predicted
Not recommended
Not recommended
Not recommended
Expected
Rat
Predicted
Not recommended
Not recommended
Not recommended
Expected

Tested
Tested

Species

Human

Dilution info

1/250 - 1/500

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Predicted
Predicted

Species

Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Human, Mouse, Rat

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

3 products for Alternative Product

Target data

Function

Ubiquitin-like protein involved in autophagy vesicles formation. Conjugation with ATG5 through a ubiquitin-like conjugating system involving also ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme, is essential for its function. The ATG12-ATG5 conjugate acts as an E3-like enzyme which is required for lipidation of ATG8 family proteins and their association to the vesicle membranes. As part of the ATG8 conjugation system with ATG5 and ATG16L1, required for recruitment of LRRK2 to stressed lysosomes and induction of LRRK2 kinase activity in response to lysosomal stress (By similarity).(Microbial infection) May act as a proviral factor. In association with ATG5, negatively regulates the innate antiviral immune response by impairing the type I IFN production pathway upon vesicular stomatitis virus (VSV) infection (PubMed:17709747). Required for the translation of incoming hepatitis C virus (HCV) RNA and, thereby, for the initiation of HCV replication, but not required once infection is established (PubMed:19666601).

Alternative names

APG12, APG12L, ATG12, APG12, APG12L, Ubiquitin-like protein ATG12, Autophagy-related protein 12, APG12-like

Recommended products

Rabbit Recombinant Monoclonal ATG12 antibody. Suitable for IHC-P, WB and reacts with Human, Mouse, Rat samples. Cited in 8 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR4800

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Storage information

Stable for 12 months at -20°C

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

ATG12 also known as Autophagy-related protein 12 plays a critical role in autophagy an essential cellular process for degrading and recycling cellular components. The molecular weight of ATG12 is around 14 kDa. It is part of the ATG12-conjugation system and is invariably expressed in various tissues with notable presence in the liver brain and heart. ATG12 forms a covalent bond with ATG5 in a ubiquitin-like conjugation pathway which is significant for this mechanism.

Biological function summary

ATG12 is vital for the formation of autophagosomes which are double-membraned vesicles that sequester cellular components for degradation. It functions as part of a complex by binding to ATG5 and further associates with ATG16L1 to form a larger ATG12-ATG5-ATG16 complex. This complex localizes to the outer membrane of the growing autophagosome facilitating elongation and closure of the vesicle making it effective in the regulation of autophagy.

Pathways

ATG12's interaction with other proteins like ATG5 places it in the macroautophagy pathway an important process maintaining cellular homeostasis and turnover of cellular components. In addition to this ATG12 also contributes to cellular responses to stress conditions via this pathway. Furthermore it has connections with signaling pathways involving proteins such as LC3 which marks autophagosomal membranes and interacts with ATG12-conjugated complexes enhancing the autophagic flux.

Associated diseases and disorders

Researchers associate dysregulated expression of ATG12 with neurodegenerative diseases such as Parkinson’s disease where impaired autophagy leads to toxic protein accumulation. Additionally alterations in ATG12 expression have links to cancer as enhanced autophagy in some tumor cells supports survival under metabolic stress. Both disorders demonstrate connections to proteins involved in the autophagy pathway suggesting therapeutic potential in modulating ATG12 activity.

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:
Terms & Conditions.

5 product images

  • Western blot - Anti-ATG12 antibody [EPR4800] (ab109491), expandable thumbnail

    Western blot - Anti-ATG12 antibody [EPR4800] (ab109491)

    False colour image of Western blot: Anti-ATG12 antibody [EPR4800] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109491 was shown to bind specifically to ATG12. A band likely to be the unfunctional complex with ATG5 was observed at 52 kDa in wild-type THP-1 cell lysates with no signal observed at this size in Atg12 knockout cell line Human ATG12 knockout THP-1 cell line ab277831 (knockout cell lysate ab278183) - unconjugated functional form not observed at 15 kDa. To generate this image, wild-type and Atg12 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-ATG12 antibody [EPR4800] (ab109491) at 1/1000 dilution

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

    Lane 2: ATG12 knockout THP-1 cell lysate at 20 µg

    Lane 3: HCT 116 cell lysate at 20 µg

    Lane 4: HeLa cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 15 kDa

    Observed band size: 52 kDa

  • Western blot - Anti-ATG12 antibody [EPR4800] (ab109491), expandable thumbnail

    Western blot - Anti-ATG12 antibody [EPR4800] (ab109491)

    Lane 1: Wild type HAP1 whole cell lysate (20 μg)
    Lane 2: ATG12 knockout HAP1 whole cell lysate (20 μg)
    Lane 3: HeLa whole cell lysate (20 μg)
    Lane 4: HCT116 whole cell lysate (20 μg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109491 observed at 55 kDa. Observed band shows ATG12-ATG5 conjugation. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

    ab109491 was shown to specifically react with

    ATG12 in wild-type cells along with additional cross-reactive bands. The band was not seen in ATG12 knockout HAP1 cells. Wild-type and ATG12 knockout samples were subjected to SDS-PAGE. ab109491 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with 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 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-ATG12 antibody [EPR4800] (ab109491)

    Predicted band size: 15 kDa

    Observed band size: 55 kDa

  • Western blot - Anti-ATG12 antibody [EPR4800] (ab109491), expandable thumbnail

    Western blot - Anti-ATG12 antibody [EPR4800] (ab109491)

    Observed band shows ATG12-ATG5 conjugation

    All lanes: Western blot - Anti-ATG12 antibody [EPR4800] (ab109491) at 1/1000 dilution

    Lane 1: Human fetal kidney tissue lysate at 10 µg

    Lane 2: HepG2 cell lysate at 10 µg

    Lane 3: Raji lysate at 10 µg

    Predicted band size: 15 kDa

  • Western blot - Anti-ATG12 antibody [EPR4800] (ab109491), expandable thumbnail

    Western blot - Anti-ATG12 antibody [EPR4800] (ab109491)

    All lanes: Western blot - Anti-ATG12 antibody [EPR4800] (ab109491) at 1/1000 dilution

    Lane 1: ATG12 transfected 293T lysate at 10 µg

    Lane 2: Non-transfected 293T lysate at 10 µg

    Predicted band size: 15 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR4800] (ab109491), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG12 antibody [EPR4800] (ab109491)

    ab109491, at 1/250 dilution, staining ATG12 in paraffin-embedded Human breast carcinoma by Immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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Product protocols

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