Overview

  • Product name
    Anti-ATG7 antibody [EPR6251]
    See all ATG7 primary antibodies
  • Description
    Rabbit monoclonal [EPR6251] to ATG7
  • 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.

  • Positive control
    • 293T, HepG2 and Jurkat whole cell lysate (ab7899), Rat spleen and kidney tissue lysates, mouse spleen and kidney tissue lysates; HT-29 and HeLa cells.
  • General notes

    We have had 1 attempt at IHC-P with ab133528 in our own lab. We observed both cytoplasmic and nuclear staining on several tissues (including human stomach, kidney and pancreatic cancer), under our experimental conditions. For IHC-P on human tissues, we would recommend using ab52472.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133528 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 1/10000 - 1/50000. Predicted molecular weight: 77 kDa.

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

ICC/IF 1/100 - 1/500.

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.

  • All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/50000 dilution (purified)

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : Mouse spleen lysate
    Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 4 : Mouse kidney lysate
    Lane 5 : Rat kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/10000 dilution (purified)

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : Rat spleen lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • 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.

  • 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.

References

This product has been referenced in:
  • Wu J  et al. Long noncoding RNA UCA1 targets miR-582-5p and contributes to the progression and drug resistance of bladder cancer cells through ATG7-mediated autophagy inhibition. Onco Targets Ther 12:495-508 (2019). Read more (PubMed: 30666128) »
  • Ju S  et al. The effect and mechanism of miR-210 in down-regulating the autophagy of lung cancer cells. Pathol Res Pract N/A:N/A (2018). Read more (PubMed: 30573163) »
See all 16 Publications for this product

Customer reviews and Q&As

1-10 of 16 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Common marmoset Tissue sections (Pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Pancreas
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Liver
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 08 2018

Application
Western blot
Sample
Rat Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Cardiomyocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Feb 08 2018

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

Abcam user community

Verified customer

Submitted Feb 01 2018

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 31 2018

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

Abcam user community

Verified customer

Submitted Jan 30 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Kidney)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 19 2017

1-10 of 16 Abreviews or Q&A

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