概述

  • 产品名称Anti-Apg7抗体[EP1759Y]
    参阅全部 Apg7 一抗
  • 描述
    兔单克隆抗体[EP1759Y] to Apg7
  • 特异性This antibody reacts with Apg7.
  • 经测试应用适用于: ICC/IF, WB, IP, Flow Cyt, IHC-Pmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Apg7 (C terminal).

  • 阳性对照
    • IHC-P: Human cervical carcinoma WB: Jurkat, HepG2, HEK293 and HAP1 cell lysate ICC/IF: HT-29 and HeLa cell lysate FC: HEK293 and HeLa cells
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody. Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative versions available:
    Anti-Apg7 antibody (Alexa Fluor® 647) [EP1759Y] (ab200876)

    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.

应用

Our Abpromise guarantee covers the use of ab52472 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/100 - 1/500.

For unpurified use at 10 µg/mL.

WB 1/100000 - 1/200000. Detects a band of approximately 70 kDa (predicted molecular weight: 78 kDa).
IP 1/30 - 1/50.
Flow Cyt 1/50 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/500.

靶标

  • 功能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.
  • 组织特异性Widely expressed, especially in kidney, liver, lymph nodes and bone marrow.
  • 序列相似性Belongs to the ATG7 family.
  • 结构域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.
  • 翻译后修饰Acetylated by EP300.
  • 细胞定位Cytoplasm. Preautophagosomal structure. Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-Apg7 antibody [EP1759Y] 图像



  • Predicted band size : 78 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Apg7 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 - ab52472 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab52472 was shown to specifically react with Apg7 when Apg7 knockout samples were used. Wild-type and ProteinX knockout samples were subjected to SDS-PAGE. ab52472 and ab8245 (loading control to Apg7) were both diluted 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded human cervical carcinoma sections labelling Apg7 with purified ab52472 at a dilution of 1/500. The secondary antibody used was ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Apg7 with purified ab52472 at 1/100. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291 anti-Tubulin (mouse mAb) followed by ab150120, AlexaFluor®594 goat anti-mouse secondary both at 1/1000. Nuclei were counterstained with DAPI (blue).

    For negative control 1, rabbit primary antibody was used followed by anti-mouse secondary antibody (ab150120). For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).

  • Flow cytometry analysis of HeLa cells labelling Apg7 (red) with purified ab52472 at dilution of 1/100. The secondary antibody used was goat anti rabbit IgG (FITC) at 1/500. Cells were fixed with 4% paraformaldehyde. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • ab52472 at 1/30 dilution immunoprecipitating Apg7 in HEK293 whole cell lysate observed at 70 KDa (lanes 1 and 2).

    Lane 1 (input): HEK293 whole cell lysate 10ug

    Lane 2 (+): ab52472 + HEK293 whole cell lysate

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52472 in HEK293 whole cell lysate

    For western blotting, ab52472 was used followed by VeriBlot for IP secondary antibody (HRP) (ab131366) as the secondary antibody at a dilution of 1/10,000.

    Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

  • All lanes : Anti-Apg7 antibody [EP1759Y] (ab52472) at 1/100000 dilution

    Lane 1 : HEK293 whole cell lysate
    Lane 2 : HepG2 whole cell lysate
    Lane 3 : Jurkat whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 78 kDa
    Observed band size : 70 kDa (why is the actual band size different from the predicted?)
    Blocking and Diluting buffer 5% NFDM/TBST
  • Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) cells labelling Apg7 with purified ab52472 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at 1/1000 dilution was used as the secondary antibody. Nuclei couterstained with DAPI (blue). 

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control. 

  • Overlay histogram showing HEK293 cells stained with unpurified ab52472 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52472, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Anti-Apg7 antibody [EP1759Y] (ab52472)参考文献

This product has been referenced in:
  • Chen Y  et al. FGFR antagonist induces protective autophagy in FGFR1-amplified breast cancer cell. Biochem Biophys Res Commun N/A:N/A (2016). WB ; Human . Read more (PubMed: 26993162) »
  • Nazarko TY  et al. Peroxisomal Atg37 binds Atg30 or palmitoyl-CoA to regulate phagophore formation during pexophagy. J Cell Biol 204:541-57 (2014). Read more (PubMed: 24535825) »

See all 6 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (Hepatocytes)
Gel Running Conditions Reduced Denaturing
Loading amount 20 µg
Specification Hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Aug 16 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293T)
Loading amount 25 µg
Specification HEK293T
Treatment siATG7
Gel Running Conditions Reduced Denaturing (8%)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Sep 06 2011

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