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  1. Link

    atg8-antibody-ab4753.pdf

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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ub-like Proteins
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Anti-ATG8 抗体 (ab4753)

  • Datasheet
  • SDS
Reviews (3)Q&A (14)References (7)

Product price, shipping and contact information

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出荷および受注について

Abpromise

保証された製品品質、優れたカスタマー・サポート。 詳細はこちら。

Western blot - Anti-ATG8 antibody (ab4753)
  • Western blot - Anti-ATG8 antibody (ab4753)

Key features and details

  • Rabbit polyclonal to ATG8
  • Suitable for: WB, ELISA
  • Reacts with: Saccharomyces cerevisiae
  • Isotype: IgG

こちらの製品もご検討ください

二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

関連製品

製品の概要

  • 製品名

    Anti-ATG8 antibody
  • 製品の詳細

    Rabbit polyclonal to ATG8
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: WB, ELISAmore details
  • 種交差性

    交差種: Saccharomyces cerevisiae
  • 免疫原

    Recombinant full length protein (S. cerevisiae).

  • 特記事項

    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.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー

    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride
  • Concentration information loading...
  • 精製度

    IgG fraction
  • 特記事項(精製)

    Purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-rabbit serum.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ub-like Proteins
    • Signal Transduction
    • Protein Trafficking
    • Vesicle Transport
    • SNAPs & SNAREs
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Microbiology
    • Organism
    • Eukaryotic Microorganism
    • Yeast
    • Saccharomyces spp
    • Cardiovascular
    • Heart
    • Autophagy
    • APG gene products
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitophagy fission and fusion
    • Cancer
    • Cell Death
    • Autophagy
    • APG gene products

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.
ELISA 1/20000 - 1/100000.

ターゲット情報

  • 関連性

    ATG8 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.
  • 細胞内局在

    Cytoplasmic
  • 参照データベース

    • Entrez Gene: 852200 Saccharomyces cerevisiae
    • SwissProt: P38182 Saccharomyces cerevisiae
    • 別名

      • Apg 8 antibody
      • APG8 antibody
      • Atg8p antibody
      • AUT7 antibody
      • CVT5 antibody
      see all

    画像

    • Western blot - Anti-ATG8 antibody (ab4753)
      Western blot - Anti-ATG8 antibody (ab4753)

      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 react

    • Western blot - Anti-ATG8 antibody (ab4753)
      Western blot - Anti-ATG8 antibody (ab4753)This image is courtesy of an anonymous abreview.
      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


      Gel run under denaturing conditions.
      Primary antibody incubated for 10 minutes at 20°C.
      Blocked using 0.5% milk for 1 minute at 20°C.
      Detection method: Western lightning chemiluminescent reagent.

      See Abreview

    プロトコール

    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    データシートおよび資料

    • Datasheet
    • SDS
  • 参考文献 (7)

    ab4753 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab4753 は 7 報の論文で使用されています。

    • Li X  et al. Dehydrin MtCAS31 promotes autophagic degradation under drought stress. Autophagy N/A:1-16 (2019). PubMed: 31362589
    • Zhou S  et al. Autophagy contributes to regulate the ROS levels and PCD progress in TMV-infected tomatoes. Plant Sci 269:12-19 (2018). WB . PubMed: 29606209
    • Jo M  et al. Yeast genetic interaction screen of human genes associated with amyotrophic lateral sclerosis: identification of MAP2K5 kinase as a potential drug target. Genome Res 27:1487-1500 (2017). PubMed: 28596290
    • Spencer B  et al. a-Synuclein interferes with the ESCRT-III complex contributing to the pathogenesis of Lewy body disease. Hum Mol Genet 25:1100-15 (2016). PubMed: 26740557
    • Li S  et al. Expression of Malus xiaojinensis IRT1 (MxIRT1) protein in transgenic yeast cells leads to degradation through autophagy in the presence of excessive iron. Yeast 32:499-517 (2015). WB . PubMed: 25871543
    • 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 . 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 . PubMed: 20174468

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 17 Abreviews or Q&A

    Western blot abreview for Anti-ATG8 antibody

    Excellent
    Abreviews
    Abreviews
    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
    Read More

    Dan Plas

    Verified customer

    投稿 Dec 10 2013

    Question

    I work with P.pastoris and need an antibody against its Atg8 which is 80% identical to S.cerevisiae atg8. Thus I wanted to ask is there a possibility for me to get an aliquot of the ab4753 antibody to test whether it is cross-reactive against P.pastoris as well.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 17 2012

    Answer

    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 am sorry I have not been able to find the the P pastoris ATG8 amino acid sequence on the protein database sites in order to check the alignment with the immunogen. If you have the sequence available, I would be pleased to check this for you.

    If the alignment is high, this will indicate the antibody is likely to detect in P pastoris. If you would like to test the antibody in P pastoris, please do not hesitate to contact me with the sequence prior to the purchase as you may be eligible for our AbTrial testing discount program. <

    Read More

    Abcam Scientific Support

    Answered on Dec 17 2012

    Question

    How many bands will it detect in western blot,one or two?

    Read More

    Abcam community

    Verified customer

    Asked on Nov 29 2012

    Answer

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

    Read More

    Abcam Scientific Support

    Answered on Nov 29 2012

    Question

    Do you think, based on sequence homology ab4753 will work with plants such as Tomato, Tobacco or Arabidopsis?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 31 2012

    Answer

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

    Read More

    Abcam Scientific Support

    Answered on Oct 31 2012

    Question

    Dear Sir or Madam,
    Thank you for the information. I am interested in the antibody but I am wondering whether it will work on Chlamydomonas.
    Isn't it possible to have a small aliquote to test?

    Best,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 24 2012

    Answer

    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 is possible that the antibody will be cross-reactive with Chlamydomonas, but we can't say for sure.
    We don't offer small samples for testing, but we do have a testing discount program if you'd still like to try this antibody. As part of this program, you would first purchase and test ab4753, and then submit an Abreview with your data and protocol. In exchange we will give you a $347 discount (the price of ab4753) on a future purchase.
    I hope that this information will be useful, but if you have further questions or need anything else, please let me know and I'll be happy to help.

    Read More

    Abcam Scientific Support

    Answered on Oct 24 2012

    Question

    Hola :
    Te reenvío nuevas imágenes y el comentario del usuario, menciona que otro lote si le funciono, ya pregunte que lote inicialmente le había funcionado. Y adjunto las imágenes del que no funciono Saludos Cordiales

    Read More

    Abcam community

    Verified customer

    Asked on Sep 25 2012

    Answer

    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 reemplazarle o reembolsarle su importe.

    El único lote actualmente disponible de este anticuerpo es el mismo que recibió el cliente en el último pedido, me temo que no hay más en stock.

    Espero tu respuesta para que me indiques cómo proceder y poder ayudar al cliente.

    Read More

    Abcam Scientific Support

    Answered on Sep 25 2012

    Question

    Hola Abcam:
    Un usuario nos reporto el siguiente problema con el anticuerpo ab4753 lote XXXX
    Ese anticuerpo fue comprado por otro distribuidor no autorizado de abcam, pero CTR se hará cargo de esta reclamación
    Gracias¡

    Read More

    Abcam community

    Verified customer

    Asked on Sep 19 2012

    Answer

    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 indica que éste reconoce otras bandas intensas correspondientes a derivados de ubiquitinas.

    Sin embargo, atendiendo al protocolo seguido, hay una serie de sugerencias que posiblemente mejoraran el rendimiento del anticuerpo, disminuyendo el ruido de fondo:

    - Usar inhibidores enzimáticos, especialmente inhibidores de proteasas, evitan la degradación de la proteína.

    -Para aumentar las condiciones reductoras yo sugeriría aumentar el tiempo de hervido de las muestras a 10 minutos.

    -La dilución usada de anticuerpo es muy baja. Nosotros recomendamos usar diluciones de entre 1/4000- 1/8000. Igualmente, aumentar la dilución del secundario (por ejemplo a 1/6000) puede disminuir el background.

    -Otra sugerencia es cambiar las condiciones de bloqueo. Usando BSA al 5% durante 2 horas.

    Confió en que estas recomendaciones mejoren el resultado. En caso contrario, por favor, no dudéis en contactarme de nuevo.

    Read More

    Abcam Scientific Support

    Answered on Sep 19 2012

    Question

    Please, could you help us with this reclamation? I have attached questionnaire of problem.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 28 2012

    Answer

    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 specific results using less primary antibody. We typically recommend using a 1:4000 dilution. It is also possible that the customer's prolonged washes are removing the blocking agent from the membrane and causing background. For that reason I would recommend performing 3 x 5 minute washes.

    Regarding the lack of signal, it may be due to the lysis in the 100% trichloroacetic acid. Using a gentler lysis buffer such as a 1% NP-40 or 1% Triton X-100 in TBS may help to better preserve the protein epitopes.

    I hope this helps to improve the customer's results. If not, please let me know the original order number and I will be happy to offer a free of charge replacement or credit.

    Read More

    Abcam Scientific Support

    Answered on Aug 28 2012

    Question

    Thanks. The Atg8 gene was removed in Atg8 mutant, thus there is not Atg8 protein expression in this mutant.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 24 2012

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on Aug 24 2012

    Question

    Thank you for your help.

    I have tried different options including lysis buffer (PBS, TBS ( same as the ref you sent to me) for glass beats assay, other commercial one), different detergent, increasing protein loading (20-30 ug) , and increase antibodies (1:5000 to 1:3000), so far I did not get any lucky. If there is an option to refund, and I would like to choose it.

    I appreciate you for your dedication to customers.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 23 2012

    Answer

    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 antibody might still be specific and just detect a band at 27 kDa ( a dimer?).

    In any case, please see below for the refund information.

    Your credit note ID for the refund is xxx.

    I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

    The credit note ID is for your reference only and does not automatically guarantee the credit.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

    Abcam Scientific Support

    Answered on Aug 23 2012

    1-10 of 17 Abreviews or Q&A

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