Recombinant
RabMAb

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

Rabbit recombinant monoclonal PKR (phospho T446) antibody [E120]. Validated in WB, IP, IHC, ChIP and tested in Human. Cited in 55 publication(s). Independently reviewed in 6 review(s).

Reviews (6)Q&A (18)References (67)

Overview

  • Product name

    Anti-PKR (phospho T446) antibody [E120]
    See all PKR primary antibodies

  • Description

    Rabbit monoclonal [E120] to PKR (phospho T446)

  • Host species

    Rabbit

  • Specificity

    The antibody only detects PKR phosphorylated on Threonine 446.

  • Tested applications

    Suitable for: IHC-P, WB, IPmore details
    Unsuitable for: Flow Cyt or ICC/IF

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse

  • Immunogen

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

  • Positive control

    • 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.

  • General notes

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

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid

  • Storage instructions

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

  • Storage buffer

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

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    E120

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab32036 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
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
WB 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IP 1/10.
  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.

    • Target

    • Function

      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.

    • Sequence similarities

      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.

    • Post-translational
      modifications

      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

    • Database links

      • Alternative names

        • 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

      Images

      • 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 1/5000 dilution

        Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates
        Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Calyculin A whole cell lysates

        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?



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

        Exposure Time: 30 seconds.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR (phospho T446) antibody [E120] (ab32036)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR (phospho T446) antibody [E120] (ab32036)

        Immunohistochemical analysis of PKR (phospho T446) expression in normal colon tissue, using 1/50 unpurified ab32036. Heat mediated antigen retrieval performed before IHC staining.

      • 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?

      • 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 (purified)

        Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) treated with Calyculin A and TNF-alpha whole cell lysates
        Lane 2 : Untreated HeLa whole cell lysates

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



        Blocking buffer: 5% NFDM/TBST

        Dilution buffer: 5% NFDM/TBST

      References

      This product has been referenced in:

      See all 67 Publications for this product

      Publishing research using ab32036? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      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

      Dr. Tamas Czompoly

      Verified customer

      Submitted 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

      Abcam user community

      Verified customer

      Submitted 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

      Abcam user community

      Verified customer

      Submitted 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

      Abcam user community

      Verified customer

      Submitted 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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