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リコンビナントRabMAb

Anti-PKR (phospho T446) 抗体 [E120] (ab32036)

Reviews (6)Q&A (18)References (120)
Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)
  • Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)
  • Immunoprecipitation - Anti-PKR (phospho T446) antibody [E120] (ab32036)
  • Anti-PKR (phospho T446) antibody [E120] (ab32036)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E120] to PKR (phospho T446)
  • Suitable for: WB, IP
  • Reacts with: Human, Pig

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-PKR (phospho T446) antibody [E120]
    PKR 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [E120] to PKR (phospho T446)

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IPmore details
    適用なし: Flow Cyt,ICC/IF or IHC-P

  • 種交差性

    交差種: Human, Pig

  • 免疫原

    Synthetic peptide within Human PKR. The exact sequence is proprietary.
    Database link: P19525
    (Peptide available as ab181660)

  • ポジティブ･コントロール

    • WB: HeLa treated with Calyculin A and TNF-alpha whole cell lysate; Calynculin A and IFN treated HeLa cell lysate. IHC-P: Human colon tissue. IP: HeLa

  • 特記事項

    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.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

  • バッファー

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • 精製度

    Protein A purified

  • ポリ/モノ

    モノクローナル

  • クローン名

    E120

  • アイソタイプ

    IgG

  • 研究分野

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab32036の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (6)
1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IP
1/10.
特記事項
WB
1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IP
1/10.
追加情報
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

ターゲット情報

  • 機能

    Following activation by double-stranded RNA in the presence of ATP, the kinase becomes autophosphorylated and can catalyze the phosphorylation of the translation initiation factor EIF2S1, which leads to an inhibition of the initiation of protein synthesis. Double-stranded RNA is generated during the course of a viral infection.

  • 配列類似性

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.
    Contains 2 DRBM (double-stranded RNA-binding) domains.
    Contains 1 protein kinase domain.

  • 翻訳後修飾

    Autophosphorylated on several Ser and Thr residues. Autophosphorylation of Thr-451 is dependent on Thr-446 and is stimulated by dsRNA binding and dimerization. Autophosphorylation apparently leads to the activation of the kinase.
  • Information by UniProt

  • 参照データベース

    • 別名

      • Double stranded RNA activated protein kinase; antibody
      • E2AK2_HUMAN antibody
      • eIF-2A protein kinase 2 antibody
      • EIF2AK1 antibody
      • EIF2AK2 antibody
      • Eukaryotic translation initiation factor 2 alpha kinase 2 antibody
      • Eukaryotic translation initiation factor 2-alpha kinase 2 antibody
      • HGNC:9437 antibody
      • Interferon induced double stranded RNA activated protein kinase antibody
      • Interferon inducible elF2 alpha kinase antibody
      • Interferon inducible RNA dependent protein kinase antibody
      • Interferon-induced, double-stranded RNA-activated protein kinase antibody
      • Interferon-inducible RNA-dependent protein kinase antibody
      • MGC126524 antibody
      • P1/eIF-2A protein kinase antibody
      • P1/eIF2A protein kinase antibody
      • p68 kinase antibody
      • PKR antibody
      • PPP1R83 antibody
      • PRKR antibody
      • Protein kinase interferon inducible double stranded RNA dependent antibody
      • Protein kinase RNA activated antibody
      • Protein kinase RNA-activated antibody
      • Protein phosphatase 1 regulatory subunit 83 antibody
      • Serine/threonine protein kinase TIK antibody
      • Tyrosine protein kinase EIF2AK2 antibody
      see all

    画像

    • Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)
      Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)
      All lanes : Anti-PKR (phospho T446) antibody [E120] (ab32036) at 1/2000 dilution

      Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
      Lane 2 : HeLa (human cervix adenocarcinoma) treated with 100 ng/mL Calyculin A whole cell lysate
      Lane 3 : HeLa (human cervix adenocarcinoma) treated with 100 ng/mL Calyculin A followed by 20 ng/mL TNF alpha whole cell lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 62 kDa
      Observed band size: 68 kDa why is the actual band size different from the predicted?


      Exposure time: 10 seconds


      Blocking/Diluting buffer: 5% NFDM/TBST

      Positive control: Rabbit monoclonal [Y117] to PKR (ab32506).

    • Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)
      Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)
      All lanes : Anti-PKR (phospho T446) antibody [E120] (ab32036) at 1/1000 dilution (unpurified)

      Lane 1 : Hela (human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : Hela cell lysate + IFN + CalyculinA
      Lane 3 : Hela cell lysate + Pptase

      Predicted band size: 62 kDa
      Observed band size: 68 kDa why is the actual band size different from the predicted?

    • Immunoprecipitation - Anti-PKR (phospho T446) antibody [E120] (ab32036)
      Immunoprecipitation - Anti-PKR (phospho T446) antibody [E120] (ab32036)

      PKR was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A(100nM 30min) whole cell lysate 10 µg with ab32036 at 1/50 dilution (2µg). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A(100nM 30min) whole cell lysate 10 µg

      Lane 2: ab32036 IP in HeLa treated with Calyculin A(100nM 30min) whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab32036 in HeLa treated with Calyculin A(100nM 30min) whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Lower bands could be cleavage form. (PMID:28702377)

    • Anti-PKR (phospho T446) antibody [E120] (ab32036)
      Anti-PKR (phospho T446) antibody [E120] (ab32036)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (120)

    ab32036 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。

    ab32036 は 120 報の論文で使用されています。

    • Peng Z  et al. PKR deficiency delays vascular aging via inhibiting GSDMD-mediated endothelial cell hyperactivation. iScience 26:105909 (2023). PubMed: 36691613
    • Zhu Y  et al. Polynucleotide phosphorylase protects against renal tubular injury via blocking mt-dsRNA-PKR-eIF2α axis. Nat Commun 14:1223 (2023). PubMed: 36869030
    • Parameswaran J  et al. Antisense, but not sense, repeat expanded RNAs activate PKR/eIF2α-dependent ISR in C9ORF72 FTD/ALS. Elife 12:N/A (2023). PubMed: 37073950
    • Krishna S  et al. Indicators of increased ER stress and UPR in aged D2-mdx and human dystrophic skeletal muscles. Front Physiol 14:1152576 (2023). PubMed: 37179835
    • Guo L  et al. Directed natural evolution generates a next-generation oncolytic virus with a high potency and safety profile. Nat Commun 14:3410 (2023). PubMed: 37296165
    View all Publications for this product

    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 24 Abreviews or Q&A

    Western blot abreview for Anti-PKR (phospho T446) antibody [E120] - ChIP Grade

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing (10 % gel)
    Loading amount
    50 µg
    Treatment
    10 ´M tunicamycin for 30 min, 1h, 2h, 3h
    Specification
    HeLa
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Tamas Czompoly

    Verified customer

    投稿 May 19 2017

    Western blot abreview for Anti-PKR (phospho T446) antibody [E120] - ChIP Grade

     Good
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HT-1080 Cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gel)
    Loading amount
    2 µg
    Specification
    HT-1080 Cells
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Feb 28 2017

    Western blot abreview for Anti-PKR (phospho T446) antibody [E120] - ChIP Grade

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (CD4+ T cell)
    Gel Running Conditions
    Reduced Denaturing (10% acrylamide)
    Loading amount
    20000 cells
    Treatment
    before or after pIC addition (10 minutes 30°C)
    Specification
    CD4+ T cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Nov 20 2015

    Western blot abreview for Anti-PKR (phospho T446) antibody [E120]

     Good
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T)
    Loading amount
    20000 cells
    Specification
    293T
    Treatment
    100ng poly(I:C) for 6 hous
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 May 09 2013

    Question

    Our customer was really happy and he got his expected results by using this replaced antibody.
    But we faced with a new problem. Our customer just contacted us and informed us that 5 Abcam products form ORDER #1174002 did not work and after several effort they did not get any results.
    All these items has been shipped all together and we probably can connect it to shipment condition by FedEx. It would be appreciated if you can help us in that regard.
    I am eagerly looking forward to hearing from you,
    Sincerely Yours
    ab130295

    ab3443

    ab42114

    ab32036

    ab83011

    Read More
    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    I have read the details you have kindly provided and have following further questions

    - What is the problem with these products. Please fill in the attached relevant questionnaire.
    - There are 8 antibodies in this order but the problem is submitted for 5. Is it true 3 others have worked or yet to be tried by customers.
    - At what temperature customer stored the products?

    Thank you very much for your cooperation. I will look forward to hearing from you soon.

    Read More

    Question

    ich habe die Hypothese, dass es im Laufe der von mir untersuchten Infektion zur Aktivierung der Kinase PKR oder/ und GCN2 kommt. Ich wollte daher fragen, ob es möglich wäre Testproben von verschiedenen Antikörpern (ab32036; ab58301; ab75836 & ab134053) zu erhalten. So dass ich zunächst schauen kann ob meine Hypthese überhaupt stimmt und wenn ja würde ich dann natürlich auch die normalen Mengen bei Ihnen bestellen.
    Es wäre nur schade, wenn meine Hypothese nicht stimmt und die Antikörper dann bei uns verkommen, weil sie sonst niemand benötigt.




    Read More
    Answer

    Vielen Dank für Ihre Anfrage.


    Leider bieten wir generell keine kostenlosen Proben zu Testzwecken an, da wir bei über 90.000Produkten gar nicht die logistischen Möglichkeiten haben, neben unserer eigentlichen Produktpallette Teströhrchen zu lagern. Hier bei Abcam verfahren wir so, dass wir einen kostenlosen Ersatz oder eine Gutschrift verschicken, falls ein Antikörper nicht so funktioniert wie auf unserem Datenblatt beschrieben, und er innerhalb der 180Tage Garantiezeit reklamiert wird.

    Allerdings bieten wir einen besonderen Test-Discount an, wenn Sie den Antikörper in einer bisher ungetesteten Applikation oder Spezies testen möchten. Falls dies auf Ihre Experimente zutrifft, melden Sie sich bitte noch einmal bei uns vom Wissenschaftlichen Dienst, damit wir Ihre Experimente besprechen können.


    Auch wenn wir keine Test-Probenversenden, möchten wir Sie jedoch einladen, unser Abreview-System zu verwenden.Das Abreview-System erlaubt es unseren Kunden, Rezensionen unserer Produkte zu veröffentlichen und dafür Punkte zu erhalten. Diese “Abpoints” können gegen Rabatte beim Kauf weiterer Produkte oder Geschenkgutscheine wie z.B. Amazon.com-Gutscheine eingetauscht werden. Sollten Sie sich dazu entscheiden, dieses Produkt zu kaufen, teilen Sie uns bitte Ihre Ergebnisse mittels eines „Abreview“ mit. Sie können die Rezension durch einen Link direkt auf unserem Produktdatenblatt abschicken.

    Unter der folgenden URL können Sie mehr über unser Abreview-System zu erfahren:

    https://www.abcam.com/abreviews

    Ich hoffe, dies hilft Ihnen weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

    Read More

    Question

    Hello,

    I had received and since tested the replacement lot (GR75663). Unfortunately, it did not detect phospho-PKR (T446) in my samples. If possible, could I have this purchase credited to my account for future orders?

    Thanks,

    Read More
    Answer

    Thank you for contacting us.

    I am sorry that the free of charge replacement has also regrettably not worked for you on this occasion. We appreciate you contacting us and this information will remain on our quality monitoring records.

    As requested I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

    Credit ID: ####

    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

    Question

    Thanks for your kindly reply, after contact this customer, she would like to have a credit note, could you please help him to create a credit note?

    We deeply appreciate your great assistance!



    Best regards

    Read More
    Answer

    Thank you for your note.

    This is to let you know that I have just contacted and asked our Account Department to raise a credit note for you - for the cost of one vial of ab32036. For your information, the internal reference note for this credit is CN20430. You can use this credit in the near future for any of the products which are in the catalogue.

    I hope this helps and if I can assist further, please do not hesitate to contact me.

    Read More

    Question

    Thanks for your kindly reply; this customer replied your question as follow:

    I used IFN-r to induce the phosphorylation at T446, and I also used the other primary antibody from “another company” to conduct the same experiment, however, the result seems better than ab32036.

    I also attached the results in this letter, please check this and help this customer to solve the problem.

    Thanks for your kindly help

    Best regards

    Read More
    Answer

    Thank you very much for your updates.

    I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

    I can offer your customer either a new vial or a credit note. Please do let me know how you wish to proceed.

    I look forward to hearing from you soon.

    Read More

    Question

    Thanks for your kindly reply, after I contacted with this customer; she indicated she was very sorry about the error information of secondary antibody, the correct content should be” goat-anti-rabbit IgG”, I’m very sorry about this mistake, therefore, could you please help this customer to solve the problem?

    Thanks for your kindly help

    Best regards

    Read More
    Answer

    Thank you for your update.

    Could you please specific and explain how this particular phosphorylation at T446 position has been induced? Has the customer used any inducer or any positive control?

    I look forward to hearing from you soon.

    Read More

    1-10 of 24 Abreviews or Q&A

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