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    pkr-phospho-t446-antibody-e120-ab32036.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases Other Kinases
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RecombinantRabMAb

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

  • Datasheet
  • SDS
Reviews (6)Q&A (18)References (73)

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Western blot - Anti-PKR (phospho T446) antibody [E120] (ab32036)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR (phospho T446) antibody [E120] (ab32036)
  • Western blot - 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: IHC-P, WB, IP
  • Reacts with: Human, Pig

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Anti-PKR antibody [EPR19374] (ab184257)
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Human PEDF ELISA Kit (SerpinF1) (ab213815)

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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 & Species

    Application Species
    IHC-P
    Pig
    WB
    Human
    See all applications and species data
  • 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

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases

Associated products

  • Alternative Versions

    • HRP Anti-PKR (phospho T446) antibody [E120] (ab195850)
    • Anti-PKR (phospho T446) antibody [E120] - BSA and Azide free (ab220801)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human PKR protein (ab71666)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Pig
WB
Human
Application Abreviews Notes
IHC-P
1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

WB (6)
1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IP
1/10.
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.
    • Target information above from: UniProt accession P19525 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 5610 Human
      • Omim: 176871 Human
      • SwissProt: P19525 Human
      • Unigene: 131431 Human
      • 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 1/2000 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).

      • 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?

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

      Protocols

      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

      Datasheets and documents

      • Datasheet
      • SDS
    • References (73)

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

      ab32036 has been referenced in 73 publications.

      • Cheng J  et al. Typical Stress Granule Proteins Interact with the 3' Untranslated Region of Enterovirus D68 To Inhibit Viral Replication. J Virol 94:N/A (2020). PubMed: 31941779
      • Wuerth JD  et al. eIF2B as a Target for Viral Evasion of PKR-Mediated Translation Inhibition. mBio 11:N/A (2020). PubMed: 32665273
      • Visser LJ  et al. Foot-and-Mouth Disease Virus Leader Protease Cleaves G3BP1 and G3BP2 and Inhibits Stress Granule Formation. J Virol 93:N/A (2019). PubMed: 30404792
      • Sauer M  et al. DHX36 prevents the accumulation of translationally inactive mRNAs with G4-structures in untranslated regions. Nat Commun 10:2421 (2019). PubMed: 31160600
      • Hood IV  et al. Crystal structure of an adenovirus virus-associated RNA. Nat Commun 10:2871 (2019). PubMed: 31253805
      View all Publications for this product

      Customer reviews and Q&As

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

      1-6 of 6 Abreviews or Q&A

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

      Good
      Abreviews
      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
      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
      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
      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

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

      Poor
      Abreviews
      Abreviews
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (Lung epithelial)
      Loading amount
      10 µg
      Specification
      Lung epithelial
      Gel Running Conditions
      Reduced Denaturing
      Blocking step
      Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
      Read More

      Abcam user community

      Verified customer

      Submitted Jul 18 2011

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

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (HeLa cell line)
      Loading amount
      20 µg
      Specification
      HeLa cell line
      Gel Running Conditions
      Reduced Denaturing (12% gel)
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Dr. Mirjam Eckert

      Verified customer

      Submitted Apr 02 2009

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