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使用敲除细胞株进行验证重组RabMAb

重组Anti-APG5L/ATG5抗体[EPR4797] (ab109490)

Submit a review Q&A (1)References (39)
Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
  • Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
  • Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
  • Flow Cytometry (Intracellular) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
  • Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
  • Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4797] to APG5L/ATG5
  • Suitable for: Flow Cyt (Intra), WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

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

  • 宿主

    Rabbit

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-Pmore details
    不适用于: IP

  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Rat

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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 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.

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab109490于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt (Intra)
1/250.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.  
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.
说明
Flow Cyt (Intra)
1/250.

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

    see all

  • 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
    • Autophagy related 5 antibody
    • hAPG5 antibody
    • Homolog of S Cerevisiae autophagy 5 antibody
    • OTTHUMP00000040507 antibody
    see all

图片

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

    Lane 1 : Wild-type THP-1 cell lysate
    Lane 2 : ATG5 knockout THP-1 cell lysate
    Lane 3 : U-87 MG cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 32 kDa
    Observed band size: 50 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-APG5L/ATG5 antibody [EPR4797] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109490 was shown to bind specifically to APG5L/ATG5. A band was observed at 50 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG5 knockout cell line ab277835 (knockout cell lysate ab290722). To generate this image, wild-type and ATG5 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  • Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)

    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 (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.

  • Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    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 (Intracellular) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    Flow Cytometry (Intracellular) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)

    Intracellular 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 Fluorr®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. 

     

     

  • Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    Western blot - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)

    ab109490, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human breast carcinoma by Immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)

    ab109490, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human kidney by Immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)
    Anti-APG5L/ATG5 antibody [EPR4797] (ab109490)

文献 (39)

发表研究结果有使用 ab109490？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab109490 被引用在 39 文献中.

  • Liang Q  et al. MicroRNA-335-5p alleviates inflammatory response, airway fibrosis, and autophagy in childhood asthma through targeted regulation of autophagy related 5. Bioengineered 13:1791-1801 (2022). PubMed: 34699311
  • Xiong W  et al. Astragaloside IV - mediated endothelial progenitor cell exosomes promote autophagy and inhibit apoptosis in hyperglycemic damaged endothelial cells via miR-21/PTEN axis. Folia Histochem Cytobiol 60:323-334 (2022). PubMed: 36504133
  • Wang X  et al. The KLF4-p62 axis prevents vascular endothelial cell injury via the mTOR/S6K pathway and autophagy in diabetic kidney disease. Endokrynol Pol 73:837-845 (2022). PubMed: 36621906
  • Kallergi E  et al. Dendritic autophagy degrades postsynaptic proteins and is required for long-term synaptic depression in mice. Nat Commun 13:680 (2022). PubMed: 35115539
  • Li RL  et al. Effect and Mechanism of LRP6 on Cardiac Myocyte Ferroptosis in Myocardial Infarction. Oxid Med Cell Longev 2021:8963987 (2021). PubMed: 34712388
View all Publications for this product

客户评价及客户问答

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Question

Would like an APG5L/ATG5 antibody that is similar to ab19130 and that will recognize the cleaved protein.

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Answer

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 found any specific information about this, but if you do have additionalinformation then I would be happy to see if one of our antibodies would react with this cleavedprotein.

I look forward to hearing from you. Please let me know if youhave any questions or if there is anything else that we can do for you.

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