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  1. Link

    atg3-antibody-epr4801-ab108251.pdf

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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ub-like Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ATG3 antibody [EPR4801] (ab108251)

  • Datasheet
  • SDS
Reviews (7) Submit a question References (14)

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Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody [EPR4801] (ab108251)
  • Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)
  • Flow Cytometry (Intracellular) - Anti-ATG3 antibody [EPR4801] (ab108251)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG3 antibody [EPR4801] (ab108251)
  • 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 (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

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Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Protein
Recombinant Human ATG3 protein (ab152071)
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Human Factor VII ELISA Kit (ab190810)

View more associated products

Overview

  • 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 (Intra), ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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 (intra): HeLa cells.
  • 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

  • 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

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4801
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ub-like Proteins
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other
    • Cardiovascular
    • Heart
    • Autophagy
    • APG gene products
    • Cancer
    • Signal transduction
    • Autophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • APG gene products
    • Cancer
    • Cell Death
    • Autophagy
    • Signal Transduction
    • Cancer
    • Cell Death
    • Autophagy
    • APG gene products

Associated products

  • Alternative Versions

    • Anti-ATG3 antibody [EPR4801] - BSA and Azide free (ab226019)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human ATG3 knockout HEK-293T cell line (ab266707)
  • KO cell lysates

    • Human ATG3 knockout HEK-293T cell lysate (ab257363)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)
  • Recombinant Protein

    • Recombinant Human ATG3 protein (ab152071)

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).
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.

Flow Cyt (Intra)
1/2000.
ICC/IF (3)
1/100 - 1/250.
Notes
WB
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).
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.

Flow Cyt (Intra)
1/2000.
ICC/IF
1/100 - 1/250.
Application notes
Is unsuitable for IP.

Target

  • 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
    modifications

    Conjugated 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.
  • Target information above from: UniProt accession Q9NT62 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 41775 Mouse
    • Unigene: 3084 Rat
    see all
  • Alternative names

    • 2610016C12Rik antibody
    • Apg 3 antibody
    • APG3 antibody
    • APG3 autophagy 3 like antibody
    • APG3 like antibody
    • APG3, S. cerevisiae, homolog of antibody
    • APG3-like antibody
    • APG3L antibody
    • Apg3p antibody
    • ATG 3 antibody
    • ATG3 antibody
    • ATG3 autophagy related 3 homolog antibody
    • ATG3 autophagy related 3 homolog (S. cerevisiae) antibody
    • ATG3_HUMAN antibody
    • Autophagy 3, S. cerevisiae, homolog of antibody
    • Autophagy Apg3p/Aut1p like antibody
    • autophagy related 3 antibody
    • Autophagy related protein 3 antibody
    • Autophagy-related protein 3 antibody
    • DKFZp564M1178 antibody
    • FLJ22125 antibody
    • hApg3 antibody
    • MGC15201 antibody
    • OTTHUMP00000214547 antibody
    • OTTHUMP00000214548 antibody
    • PC3 96 antibody
    • Protein PC3-96 antibody
    • Ubiquitin-like-conjugating enzyme ATG3 antibody
    see all

Images

  • Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody [EPR4801] (ab108251)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG3 antibody [EPR4801] (ab108251)
    Immunofluorescent staining of HeLa cells using ab108251 at 1/100.
  • Western blot - Anti-ATG3 antibody [EPR4801] (ab108251)
    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 (Intracellular) - Anti-ATG3 antibody [EPR4801] (ab108251)
    Flow Cytometry (Intracellular) - Anti-ATG3 antibody [EPR4801] (ab108251)

    Intracellular 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 Fluorr® 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. 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG3 antibody [EPR4801] (ab108251)
    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.

  • Anti-ATG3 antibody [EPR4801] (ab108251)
    Anti-ATG3 antibody [EPR4801] (ab108251)

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (14)

Publishing research using ab108251? Please let us know so that we can cite the reference in this datasheet.

ab108251 has been referenced in 14 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
  • Zhang Z  et al. RNA-binding protein ZFP36/TTP protects against ferroptosis by regulating autophagy signaling pathway in hepatic stellate cells. Autophagy 16:1482-1505 (2020). PubMed: 31679460
  • He C  et al. Yunvjian-Medicated Serum Protects INS-1 Cells against Glucolipotoxicity-Induced Apoptosis through Autophagic Flux Modulation. Evid Based Complement Alternat Med 2020:8878259 (2020). PubMed: 33414841
  • 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
View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATG3 antibody [EPR4801]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293)
Permeabilization
Yes - 0.01% TritonX-100
Specification
HEK293
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 03 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATG3 antibody [EPR4801]

Average
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (H9c2 rat cardiomyocytes)
Permeabilization
Yes - 0.01% TritonX-100
Specification
H9c2 rat cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 18 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-ATG3 antibody [EPR4801]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Cardiomyocytes)
Permeabilization
Yes - 0.01% TritonX-100
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 18 2020

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