Recombinant
RabMAb

Anti-Apg7 antibody [EPR20384] (ab183188)

Overview

  • Product name
    Anti-Apg7 antibody [EPR20384]
    See all Apg7 primary antibodies
  • Description
    Rabbit monoclonal [EPR20384] to Apg7
  • Tested applications
    Suitable for: WB, ICC/IF, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Apg7 aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: O95352

  • Positive control
    • WB: Human fetal heart, kidney and liver lysates; rat liver, brain and spleen lysates; mouse brain, kidney and spleen lysates; MCF7, HEK-293, HepG2, WEHI-231, Neuro-2a, RAW 264.7, NIH/3T3 and C6 whole cell lysates. IHC: Human pancreas, mouse testis and rat panreas tissue. ICC/IF: MCF7, NIH/3TS and C6 cells. Flow Cytometry: NIH/3T3 and C6 cells. IP: Mouse liver lysate.
  • 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.

Applications

Our Abpromise guarantee covers the use of ab183188 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 1/1000. Detects a band of approximately 77 kDa (predicted molecular weight: 77 kDa).
ICC/IF 1/250.
IP 1/120.
Flow Cyt 1/250.
IHC-P 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

  • Apg7 was immunoprecipitated from 0.35 mg of mouse liver lysate with ab183188 at 1/120 dilution. Western blot was performed from the immunoprecipitate using ab183188 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: Mouse liver lysate10 µg (Input).

    Lane 2: ab183188 IP in mouse liver lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab183188 in mouse liver lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.

  • Flow cytometric analysis of 80% methanol-fixed C6 (rat glial tumor cell line) cell line labeling Apg7 with ab183188 at 1/250 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Flow cytometric analysis of 80% methanol-fixed NIH/3T3 (mouse embyro fibroblast cell line) cell line labeling Apg7 with ab183188 at 1/250 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Immunofluorescent analysis of 100% methanol-fixed C6 (rat glial tumor cell line) cells labeling Apg7 with ab183188 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on C6 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 100% methanol-fixed NIH/3T3 (mouse embyro fibroblast cell line) cells labeling Apg7 with ab183188 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 100% methanol-fixed MCF7 (human breast adenocarcinoma cell line) cells labeling Apg7 with ab183188 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF7 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling Apg7 with ab183188 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on acinus of rat pancreas is observed (PMID: 22562096). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemical analysis of paraffin-embedded mouse testes tissue labeling Apg7 with ab183188 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on mouse testis is observed (PMID: 22848486). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Apg7 with ab183188 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on acinus of human pancreas is observed (PMID: 22562096). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Anti-Apg7 antibody [EPR20384] (ab183188) at 1/1000 dilution + NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

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

    Blocking/dilution buffer: 5% NFDM/TBST

    Exposure time: 15 seconds

  • All lanes : Anti-Apg7 antibody [EPR20384] (ab183188) at 1/1000 dilution

    Lane 1 : Rat brain tissue lysate
    Lane 2 : Rat spleen tissue lysate
    Lane 3 : Mouse brain tissue lysate
    Lane 4 : Mouse kidney tissue lysate
    Lane 5 : Mouse spleen tissue lysate
    Lane 6 : C6 (rat glial tumor cell line) whole cell lysate
    Lane 7 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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

    Blocking/Dilution buffer: 5% NFDM/TBST

    Exposure time: Lane 1-6: 3 minutes; Lane 7: 30 seconds

  • All lanes : Anti-Apg7 antibody [EPR20384] (ab183188) at 1/1000 dilution

    Lane 1 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
    Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
    Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
    Lane 4 : WEHI-231 (mouse lymphoblast B cell lymphoma cell line) whole cell lysate at 20 µg
    Lane 5 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 20 µg
    Lane 6 : Rat liver tissue lysate at 20 µg
    Lane 7 : Human fetal liver tissue lysate

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 77 kDa

    Blocking/Dilution buffer: 5% NFDM/TBST

    Exposure time: 3 minutes

  • All lanes : Anti-Apg7 antibody [EPR20384] (ab183188) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

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

    Blocking/Dilution buffer: 5% NFDM/TBST

    Exposure time: 3 minutes

References

ab183188 has not yet been referenced specifically in any publications.

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

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