Overview

  • Product nameAnti-ATG8 antibody
  • Description
    Rabbit polyclonal to ATG8
  • Tested applicationsSuitable for: WB, IHC-P, ELISA, Electron Microscopymore details
  • Species reactivity
    Reacts with: Saccharomyces cerevisiae
  • Immunogen

    Recombinant full length protein (S. cerevisiae).

Properties

Applications

Our Abpromise guarantee covers the use of ab4753 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 1/4000 - 1/8000. Detects a band of approximately 14 kDa. This antibody using the specified conditions may recognize other prominent intrinsic bands (UBLs or their conjugates). Other intrinsic bands are readily detectable in yeast lysates at lower antibody dilutions.
IHC-P Use at an assay dependent concentration. PubMed: 20174468
ELISA 1/20000 - 1/100000.
Electron Microscopy Use at an assay dependent concentration. PubMed: 25195618

Target

  • RelevanceATG8 is a ubiquitin-like protein in yeast required for autophagy (intracellular bulk protein degradation). Starved yeast cells take up their own cytoplasm into vacuoles through autophagic bodies. ATG8 is requiered for autophagy: modified by the serial action of ATG4, ATG7, and ATG3, and conjugated at the C terminus with phosphatidylethanolamine, to become the form essential for generation of autophagosomes. ATG8 interacts also with the endoplasmic reticulum to Golgi v-SNARE protein BET1 and the vacuolar v-SNARE protein NYV1. ATG8 is highly conserved, with apparent homologues in the worm, mammals and plants. In higher eukaryotes, ATG8 consists of a multigene family.
  • Cellular localizationCytoplasmic
  • Database links
    • Alternative names
      • Apg 8 antibody
      • APG8 antibody
      • Atg8p antibody
      • AUT7 antibody
      • CVT5 antibody
      see all

    Anti-ATG8 antibody images

    • Ab4753, generated by immunization with recombinant yeast ATG8 (or Apg 8), was tested by immunoblot with other anti-UBL (Ubiquitin-like modifier) antibodies against E.coli lysates expressing the ATG8-GFP fusion protein (Apg 8-GFP in both panels)). All UBLs possess limited homology to Ubiquitin and to each other, therefore it is important to know the degree of reactivity of each antibody against each UBL.

      Panel A shows total protein staining using ponceau.

      Panel B shows specific reaction with ATG8 (Apg 8) using a 1:4,000 and 1:8,000 dilution of ab4753 followed by reaction with a 1:15,000 dilution of HRP Goat-a-Rabbit IgG (ab7090). All primary antibodies were diluted in TTBS buffer supplemented with 5% non-fat milk and incubated with the membranes overnight at 4° C. E.coli lysate proteins were separated by SDS-PAGE using a 15% gel.

      Similar experiments (data not shown), where other UBL fusion proteins were separated and probed with this antibody showed no reactivity of anti-APG8 (anti-Apg 8) with other UBLs. This data indicates that anti-ATG8 (anti-Apg8)is highly specific and does not cross react with other UBLs. A chemiluminescence system was used for signal detection (Roche). Other detection systems will yield similar results.

      Data contributed by M. Malakhov, www.lifesensors.com, personal communication.

    • Anti-ATG8 antibody (ab4753) at 1/1500 dilution + Whole cell lysate prepared from Human huh-7 cells expressing the Saccharomyces cerevisiae protein at 15 µg

      Secondary
      Sheep anti-rabbit IgG conjugated to HRP at 1/7000 dilution

      Observed band size : 15 kDa (why is the actual band size different from the predicted?)


      Exposure time : 4 minutes

      This image is courtesy of an anonymous abreview.

      See Abreview

    References for Anti-ATG8 antibody (ab4753)

    This product has been referenced in:
    • Schatz D  et al. Hijacking of an autophagy-like process is critical for the life cycle of a DNA virus infecting oceanic algal blooms. New Phytol 204:854-63 (2014). WB, Electron Microscopy . Read more (PubMed: 25195618) »
    • Crews L  et al. Selective Molecular Alterations in the Autophagy Pathway in Patients with Lewy Body Disease and in Models of alpha-Synucleinopathy. PLoS One 5:e9313 (2010). WB, IHC-P ; Human, Mouse . Read more (PubMed: 20174468) »

    See all 2 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 35 µg
    Gel Running Conditions Non-reduced Non-Denaturing (Native)
    Sample Lymnaea stagnalis (Great Pond Snail) Cell lysate - whole cell (CNS (circumesophogeal ganglia))
    Specification CNS (circumesophogeal ganglia)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
    Username

    Dan Plas

    Verified customer

    Submitted Dec 10 2013

    As far as we are aware, ab4753 has never been tested with samples from P.pastoris. All tested species by our 6 month guarantee are stated on our datasheets, and these are updated as soon as any new information is brought to our attention.

    I ...

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    This antibody will primarily detect one band at 14 kDa, but may also recognize other prominent intrinsic bands corresponding to UBLs or their conjugates.

    Could you send us the protein sequences specific to species in question?We can then check for homology.

    Thank you very much for contacting us with your question.
    I ran an alignment between the immunogen of ab4753 (full-length S. cerevisiae protein) and the C. reinhardtii autophagy protein sequence, and there is 75% identity between the sequences. It...

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    Muchas gracias por tu respuesta y por enviar las imágenes.

    Efectivamente es posible que el vial concreto que hemos enviado al cliente se haya degradado. Dado que el producto está bajo garantía, estaré encantada de ...

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    Gracias por contactarnos y mandarnos el cuestionario completo. Lamentablemente no he recibido la imagen adjunta en el cuestionario, por lo que agradecería que si es posible la volvierais a enviar.

    En la datasheet del anticuerpo se indi...

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    Thank you for contacting us. Unfortunately, I was unable to view the image provided in the word document. Is is possible for you to send a copy of the image separately?

    Based on the customer's protocol, it may be possible to obtain more speci...

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    Thank you for your email.

    I see. Yes, in that case the antibody is indeed not working correctly. I apologize about this.

    I wish you good luck with your research, and please let me know if I can help you with anything else.

    Thank you for your reply. I'm sorry that the antibody did not work. I was wondering if the ATG8 mutant you were refering to is a deletion mutant (ATG8 protein is absent) or just point or fragmentmutation in the ATG8 protein? In case of the latter, the ...

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    1-10 of 17 Abreviews or Q&A

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