Recombinant
RabMAb

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

Abreviews (6)Q&A (18)Specific References (49)

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: ChIP, 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.
    (Peptide available as ab181660)

  • Positive control
    • Calynculin A and TNF alpha treated HeLa cell lysate
  • 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.

    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.

    This product is a recombinant rabbit monoclonal antibody.

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
ChIP Use a concentration of 4 µg/ml.
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).Can be blocked with PKR (phospho T446) peptide (ab181660).
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 1/1000 dilution (purified)

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

      • 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 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?)

      • 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.

      References

      This product has been referenced in:
      • Krishnamoorthy J  et al. Downregulation of PERK activity and eIF2a serine 51 phosphorylation by mTOR complex 1 elicits pro-oxidant and pro-death effects in tuberous sclerosis-deficient cells. Cell Death Dis 9:254 (2018). WB . Read more (PubMed: 29449538) »
      • Tongtako W  et al. Canine dorsal root ganglia satellite glial cells represent an exceptional cell population with astrocytic and oligodendrocytic properties. Sci Rep 7:13915 (2017). Dog . Read more (PubMed: 29066783) »

      See all 49 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

      Submit a review Submit a question

      24 reviews or Q&A

      10
      Votes: Highest First

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

       Good
      Abreviews
      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
      abreview image
      Username

      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
      Username

      Abcam user community

      Verified customer

      Submitted Feb 28 2017

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

       Good
      Abreviews
      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
      abreview image
      Username

      Abcam user community

      Verified customer

      Submitted Nov 20 2015

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

       Good
      Abreviews
      Abcam guarantees this product to work in the species/application used in this Abreview.
      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
      Username

      Abcam user community

      Verified customer

      Submitted May 09 2013

      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...

      Read More

      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...

      Read More

      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; ab75...

      Read More

      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...

      Read More

      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,

      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.
      <...

      Read More

      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

      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 cred...

      Read More

      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, ho...

      Read More

      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 th...

      Read More

      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 mi...

      Read More

      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.

      1-10 of 24 Abreviews or Q&A

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

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