Recombinant
RabMAb

Anti-ATG3 antibody [EPR4801] (ab108251)

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: IHC-Fr, WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ATG3 (N terminal). The exact sequence is proprietary.

  • Positive control
    • WB: K562, HL-60, HeLa or Jurkat cell lysates. IHC-P: Human muscle tissue. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
  • General 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 rabbit monoclonal antibody.

Properties

Applications

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
IHC-Fr Use at an assay dependent concentration.
WB 1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).
IHC-P 1/100 - 1/250. Perform antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 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.
    • Information by UniProt
    • Database links
    • 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

    • 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?)

    • ab108251 staining ATG3 in Mouse uterus tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde and blocked with 5% BSA for 1 hour at 25°C. Samples were incubated with primary antibody (1/200 in 1% BSA in PBS) for 12 hours at 4°C. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

      See Abreview

    • Immunofluorescent staining of HeLa cells using ab108251 at 1/100.
    • 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.

    • ab108251, at a 1/100 dilution, staining Human ATG3 in muscle, using Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue.

    References

    This product has been referenced in:
    • Puri C  et al. The RAB11A-Positive Compartment Is a Primary Platform for Autophagosome Assembly Mediated by WIPI2 Recognition of PI3P-RAB11A. Dev Cell 45:114-131.e8 (2018). ICC/IF . Read more (PubMed: 29634932) »
    • Russ DW  et al. Muscle-specificity of age-related changes in markers of autophagy and sphingolipid metabolism. Biogerontology 16:747-59 (2015). WB ; Rat . Read more (PubMed: 26296420) »

    See all 3 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Lung)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    15 µg
    Treatment
    1x10^7 S. aureus peritoneal clot
    Specification
    Lung
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Mr. Alan Chang

    Verified customer

    Submitted Mar 20 2014

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (uterus)
    Permeabilization
    No
    Specification
    uterus
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Formaldehyde
    Username

    Dr. Varkha Agrawal

    Verified customer

    Submitted Nov 14 2013

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (placenta)
    Gel Running Conditions
    Reduced Denaturing (4-12)
    Loading amount
    5 µg
    Specification
    placenta
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Varkha Agrawal

    Verified customer

    Submitted Nov 14 2013

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Uterus)
    Permeabilization
    No
    Specification
    Uterus
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Username

    Dr. Mukesh Jaiswal

    Verified customer

    Submitted Nov 13 2013

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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