使用敲除细胞株进行验证RabMAb

Anti-APG5L/ATG5抗体[EPR4797] (ab109490)

概述

  • 产品名称Anti-APG5L/ATG5抗体[EPR4797]
    参阅全部 APG5L/ATG5 一抗
  • 描述
    兔单克隆抗体[EPR4797] to APG5L/ATG5
  • 经测试应用适用于: Flow Cyt, WB, IHC-P, ICCmore details
    不适用于: IP
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    corresponding to Human APG5L/ATG5 (N terminal).

  • 阳性对照
    • THP1, Raji, HeLa, HT1080 and Human fetal kidney lysates; Human breast carcinoma and Human kidney tissues.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    Alternative versions available:

    Anti-APG5L/ATG5 antibody (Alexa Fluor® 488) [EPR4797] (ab206714)
    Anti-APG5L/ATG5 antibody (Alexa Fluor® 647) [EPR4797] (ab206715)

     

    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.

应用

Our Abpromise guarantee covers the use of ab109490 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
Flow Cyt 1/250.
WB 1/1000 - 1/10000. Predicted molecular weight: 32 kDa.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC 1/100 - 1/250.
  • 应用说明Is unsuitable for IP.
  • 靶标

    • 功能Involved in autophagic vesicle formation. Conjugation with ATG12, through a ubiquitin-like conjugating system involving ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme, is essential for its function. The ATG12-ATG5 conjugate acts as an E3-like enzyme which is required for lipidation of ATG8 family proteins and their association to the vesicle membranes. Involved in mitochondrial quality control after oxidative damage, and in subsequent cellular longevity. The ATG12-ATG5 conjugate also negatively regulates the innate antiviral immune response by blocking the type I IFN production pathway through direct association with RARRES3 and MAVS. Also plays a role in translation or delivery of incoming viral RNA to the translation apparatus. Plays a critical role in multiple aspects of lymphocyte development and is essential for both B and T lymphocyte survival and proliferation. Required for optimal processing and presentation of antigens for MHC II. Involved in the maintenance of axon morphology and membrane structures, as well as in normal adipocyte differentiation. Promotes primary ciliogenesis through removal of OFD1 from centriolar satellites and degradation of IFT20 via the autophagic pathway.
      May play an important role in the apoptotic process, possibly within the modified cytoskeleton. Its expression is a relatively late event in the apoptotic process, occurring downstream of caspase activity. Plays a crucial role in IFN-gamma-induced autophagic cell death by interacting with FADD.
    • 组织特异性Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells.
    • 序列相似性Belongs to the ATG5 family.
    • 翻译后修饰Conjugated to ATG12; which is essential for autophagy, but is not required for association with isolation membrane.
      Acetylated by EP300.
    • 细胞定位Cytoplasm. Preautophagosomal structure membrane. Colocalizes with nonmuscle actin. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed (By similarity). 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
      • APG 5 antibody
      • APG 5L antibody
      • APG5 antibody
      • APG5 autophagy 5 like antibody
      • APG5 like antibody
      • APG5-like antibody
      • APG5L antibody
      • Apoptosis specific protein antibody
      • Apoptosis-specific protein antibody
      • ASP antibody
      • ATG 5 antibody
      • Atg5 antibody
      • ATG5 autophagy related 5 homolog antibody
      • ATG5_HUMAN antibody
      • Autophagy protein 5 antibody
      • hAPG5 antibody
      • Homolog of S Cerevisiae autophagy 5 antibody
      • OTTHUMP00000040507 antibody
      see all

    Anti-APG5L/ATG5 antibody [EPR4797] 图像



    • Predicted band size : 32 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: APG5L/ATG5 knockout HAP1 cell lysate (20 µg)
      Lane 3: Raji cell lysate (20 µg)
      Lane 4: Jeg-3 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab109490 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
      ab109490 was shown to specifically react with APG5L/ATG5 when APG5L/ATG5 knockout samples were used. Wild-type and APG5L/ATG5 knockout samples were subjected to SDS-PAGE. ab109490 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • All lanes : Anti-APG5L/ATG5 antibody [EPR4797] (ab109490) at 1/1000 dilution

      Lane 1 : THP1 cell lysate
      Lane 2 : Raji cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : HT1080 cell lysate

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 32 kDa
    • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling APG5L/ATG5 with purified ab109490 at 1/250 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    • Anti-APG5L/ATG5 antibody [EPR4797] (ab109490) at 1/1000 dilution + Human fetal kidney lysate at 10 µg

      Predicted band size : 32 kDa
      Upper band shows APG5L/ATG5 conjugated to ATG12.
    • ab109490, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human breast carcinoma by Immunohistochemistry.
    • ab109490, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human kidney by Immunohistochemistry.

    Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)参考文献

    This product has been referenced in:
    • He G  et al. Gadd45b prevents autophagy and apoptosis against rat cerebral neuron oxygen-glucose deprivation/reperfusion injury. Apoptosis 21:390-403 (2016). WB ; Rat . Read more (PubMed: 26882903) »
    • Pla A  et al. TLR4 mediates the impairment of ubiquitin-proteasome and autophagy-lysosome pathways induced by ethanol treatment in brain. Cell Death Dis 5:e1066 (2014). WB ; Mouse . Read more (PubMed: 24556681) »

    See all 2 Publications for this product

    Product Wall

    Thank you for your call today and for your questions about our APG5L/ATG5 antibodies.

    Have you found any further information regarding the sequence of the cleaved protein (e.g. whether it is an N-terminal or C-terminal fragment)? I have not f...

    Read More

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