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Cell Biology Apoptosis Intracellular Associated Proteins
使用敲除细胞株进行验证重组RabMAb

重组Anti-ATG7抗体[EP1759Y] (ab52472)

  • Datasheet
  • SDS
Reviews (3) Submit a question References (35)

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Western blot - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Flow Cytometry - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Immunoprecipitation - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Western blot - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Flow Cytometry - Anti-ATG7 antibody [EP1759Y] (ab52472)
  • Anti-ATG7 antibody [EP1759Y] (ab52472)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1759Y] to ATG7
  • Suitable for: ICC/IF, WB, IP, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Human

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概述

  • 产品名称

    Anti-ATG7抗体[EP1759Y]
    参阅全部 ATG7 一抗
  • 描述

    兔单克隆抗体[EP1759Y] to ATG7
  • 宿主

    Rabbit
  • 特异性

    This antibody reacts with ATG7.
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data
  • 免疫原

    Synthetic peptide within Human ATG7 (C terminal). The exact sequence is proprietary.
    Database link: O95352

  • 阳性对照

    • IHC-P: Human cervical carcinomaWB: Jurkat, HepG2, HEK293 and HAP1 cell lysate ICC/IF: HT-29 and HeLa whole cell lysate (ab150035)Flow Cyt: HEK293 and HeLa cells
  • 常规说明

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

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.21% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    EP1759Y
  • 同种型

    IgG
  • 研究领域

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Associated Proteins
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ub-like Proteins
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cardiovascular
    • Heart
    • Autophagy
    • APG gene products
    • Cancer
    • Signal transduction
    • Autophagy
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitophagy fission and fusion
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • APG gene products
    • Metabolism
    • Types of disease
    • Cancer
    • Cancer
    • Cell Death
    • Autophagy
    • Signal Transduction
    • Cancer
    • Cell Death
    • Autophagy
    • APG gene products

相关产品

  • Alternative Versions

    • Alexa Fluor® 488 Anti-ATG7 antibody [EP1759Y] (ab201251)
    • PE Anti-ATG7 antibody [EP1759Y] (ab211584)
    • Anti-ATG7 antibody [EP1759Y] - BSA and Azide free (ab227564)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat cytoplasmic extract lysate (ab14842)

应用

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Human
应用 Ab评论 说明
ICC/IF
1/100 - 1/500.

For unpurified use at 10 µg/mL.

WB (2)
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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession O95352 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 10533 Human
    • Omim: 608760 Human
    • SwissProt: O95352 Human
    • Unigene: 38032 Human
    • 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

    图片

    • Western blot - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Western blot - Anti-ATG7 antibody [EP1759Y] (ab52472)

      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 - ab52472 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab52472 was shown to specifically react with ATG7 when ATG7 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG7 antibody [EP1759Y] (ab52472)

      Immunohistochemical analysis of paraffin-embedded human cervical carcinoma sections labelling ATG7 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 - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EP1759Y] (ab52472)

      Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ATG7 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 - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Flow Cytometry - Anti-ATG7 antibody [EP1759Y] (ab52472)

      Flow cytometry analysis of HeLa cells labelling ATG7 (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.

    • Immunoprecipitation - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Immunoprecipitation - Anti-ATG7 antibody [EP1759Y] (ab52472)

      ab52472 at 1/30 dilution immunoprecipitating ATG7 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 Detection Reagent (HRP) (ab131366) for detection at a dilution of 1/10,000.

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

    • Western blot - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Western blot - Anti-ATG7 antibody [EP1759Y] (ab52472)
      All lanes : Anti-ATG7 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
      All lanes : 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 - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EP1759Y] (ab52472)

      Immunocytochemistry/Immunofluorescence analysis of HT-29 (human colorectal adenocarcinoma) cells labelling ATG7 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. 

    • Flow Cytometry - Anti-ATG7 antibody [EP1759Y] (ab52472)
      Flow Cytometry - Anti-ATG7 antibody [EP1759Y] (ab52472)

      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-ATG7 antibody [EP1759Y] (ab52472)
      Anti-ATG7 antibody [EP1759Y] (ab52472)

    实验方案

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (35)

    发表研究结果有使用 ab52472?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab52472 被引用在 35 文献中.

    • Ma Z  et al. PHLDA2 regulates EMT and autophagy in colorectal cancer via the PI3K/AKT signaling pathway. Aging (Albany NY) 12:7985-8000 (2020). PubMed: 32385195
    • Li G  et al. Long non-coding RNA growth arrest-specific 5 (GAS5) acts as a tumor suppressor by promoting autophagy in breast cancer. Mol Med Rep 22:2460-2468 (2020). PubMed: 32705220
    • Yeom J  et al. Oxyresveratrol Induces Autophagy via the ER Stress Signaling Pathway, and Oxyresveratrol-Induced Autophagy Stimulates MUC2 Synthesis in Human Goblet Cells. Antioxidants (Basel) 9:N/A (2020). PubMed: 32150901
    • Xiao X  et al. LncRNA ENST00000453774.1 contributes to oxidative stress defense dependent on autophagy mediation to reduce extracellular matrix and alleviate renal fibrosis. J Cell Physiol 234:9130-9143 (2019). PubMed: 30317629
    • Feng L  et al. cis-Khellactone Inhibited the Proinflammatory Macrophages via Promoting Autophagy to Ameliorate Imiquimod-Induced Psoriasis. J Invest Dermatol 139:1946-1956.e3 (2019). PubMed: 30878677
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-3 of 3 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ATG7 antibody [EP1759Y]

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

    Mr. Carl Hobbs

    Verified customer

    提交于 Jul 26 2019

    Western blot abreview for Anti-Apg7 antibody [EP1759Y]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    提交于 Aug 16 2016

    Western blot abreview for Anti-Apg7 antibody [EP1759Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    提交于 Sep 06 2011

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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