Validated using a knockout cell lineRecombinantRabMAb

Recombinant HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)

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Western blot - HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
  • Western blot - HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
  • HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • HRP Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Conjugation: HRP

Overview

  • Product name

    HRP Anti-APG5L/ATG5 antibody [EPR1755(2)]
    See all APG5L/ATG5 primary antibodies

  • Description

    HRP Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5

  • Host species

    Rabbit

  • Conjugation

    HRP

  • Tested applications

    Suitable for: WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human APG5L/ATG5 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • WB: Raji, Raw264.7, NIH3T3 andPC-12 whole cell lysates.

  • General notes

    Alternative versions available:
    Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) - Knockout validated

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab199560 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 1/5000. Detects a band of approximately 50 kDa (predicted molecular weight: 32 kDa).

Target

  • Function

    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.

  • Tissue specificity

    Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells.

  • Sequence similarities

    Belongs to the ATG5 family.

  • Post-translational
    modifications

    Conjugated to ATG12; which is essential for autophagy, but is not required for association with isolation membrane.
    Acetylated by EP300.

  • Cellular localization

    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

  • Database links

    see all

  • Alternative names

    • 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

Images

  • Western blot - HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
    Western blot - HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
    All lanes : HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : APG5L/ATG5 knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 32 kDa
    Observed band size: 52 kDa
    why is the actual band size different from the predicted?


    Exposure time: 8 minutes


    ab199560 was shown to specifically react with ATG5 in wild-type HAP1 cells as signal was lost in ATG5 knockout cells. Wild-type and ATG5 knockout samples were subjected to SDS-PAGE. Ab199560 was incubated overnight at 4°C at a 1/5000 dilution.

  • Western blot - HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
    Western blot - HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
    All lanes : HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560) at 1/5000 dilution

    Lane 1 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 2 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 3 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab199560 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)
    HRP Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab199560)

References (0)

Publishing research using ab199560? Please let us know so that we can cite the reference in this datasheet.

ab199560 has not yet been referenced specifically in any publications.

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