Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-ATG7 antibody [EPR6251] - BSA and Azide free (ab232348)

Overview

  • Product name

    Anti-ATG7 antibody [EPR6251] - BSA and Azide free
    See all ATG7 primary antibodies
  • Description

    Rabbit monoclonal [EPR6251] to ATG7 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ATG7 aa 1-100. The exact sequence is proprietary.
    Database link: O95352

  • Positive control

    • WB: Wild-type HAP1 cell lysate; Jurkat and HepG2 cell lysates.
  • General notes

    Ab232348 is the carrier-free version of ab133528. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab232348 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Applications

Our Abpromise guarantee covers the use of ab232348 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 Use at an assay dependent concentration. Predicted molecular weight: 77 kDa.

Use 5% non-fat dry milk + TBST for blocking.

ICC/IF Use at an assay dependent concentration.

Target

  • Function

    E1-like activating enzyme involved in the 2 ubiquitin-like systems required for cytoplasm to vacuole transport (Cvt) and autophagy. Activates ATG12 for its conjugation with ATG5 as well as the ATG8 family proteins for their conjugation with phosphatidylethanolamine. Both systems are needed for the ATG8 association to Cvt vesicles and autophagosomes membranes. Required for autophagic death induced by caspase-8 inhibition. Required for mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Modulates p53/TP53 activity to regulate cell cycle and survival during metabolic stress. Plays also a key role in the maintenance of axonal homeostasis, the prevention of axonal degeneration, the maintenance of hematopoietic stem cells, the formation of Paneth cell granules, as well as in adipose differentiation.
  • Tissue specificity

    Widely expressed, especially in kidney, liver, lymph nodes and bone marrow.
  • Sequence similarities

    Belongs to the ATG7 family.
  • Domain

    The C-terminal part of the protein is essential for the dimerization and interaction with ATG3 and ATG12.
    The N-terminal FAP motif (residues 15 to 17) is essential for the formation of the ATG89-PE and ATG5-ATG12 conjugates.
  • Post-translational
    modifications

    Acetylated by EP300.
  • Cellular localization

    Cytoplasm. Preautophagosomal structure. Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme.
  • Information by UniProt
  • Database links

  • Alternative names

    • 1810013K23Rik antibody
    • Apg 7 antibody
    • APG7 autophagy 7 like antibody
    • APG7 autophagy 7-like (S. cerevisiae) antibody
    • APG7 like antibody
    • APG7, S. cerevisiae, homolog of antibody
    • APG7-like antibody
    • APG7L antibody
    • ATG 7 antibody
    • ATG12-activating enzyme E1 ATG7 antibody
    • ATG7 antibody
    • ATG7 autophagy related 7 homolog (S. cerevisiae) antibody
    • ATG7 autophagy related 7 homolog antibody
    • ATG7_HUMAN antibody
    • Atg7l antibody
    • Autophagy 7, S. cerevisiae, homolog of antibody
    • Autophagy related protein 7 antibody
    • Autophagy-related 7 (yeast) antibody
    • Autophagy-related protein 7 antibody
    • DKFZp434N0735 antibody
    • GSA 7 antibody
    • GSA7 antibody
    • hAGP7 antibody
    • Ubiquitin activating enzyme E1 like protein antibody
    • Ubiquitin-activating enzyme E1-like protein antibody
    • Ubiquitin-like modifier-activating enzyme ATG7 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATG7 with purified ab133528 at 1/150 dilution (8.5μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133528).

  • Immunocytochemistry/Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma cell line) labeling ATG7 with purified ab133528 at 1/500 dilution. Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133528).

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ATG7 knockout HAP1 cell lysate (20 µg)
    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab133528 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab133528 was shown to specifically react with ATG7 when ATG7 knockout samples were used. Wild-type and Apg7 knockout samples were subjected to SDS-PAGE. ab133528 and ab8245 (loading control to GAPDH) were diluted 1/10,000 and 1/2000 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 1/10 000 dilution for 1 h at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133528).

References

ab232348 has not yet been referenced specifically in any publications.

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