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    pkr-antibody-y117-bsa-and-azide-free-ab239817.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases Other Kinases
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
  • Flow Cytometry (Intracellular) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
  • Immunoprecipitation - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
  • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
  • Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y117] to PKR - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), IP, IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

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Recombinant human PKR protein (ab71666)

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Overview

  • Product name

    Anti-PKR antibody [Y117] - BSA and Azide free
    See all PKR primary antibodies
  • Description

    Rabbit monoclonal [Y117] to PKR - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), IP, IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • General notes

    ab239817 is the carrier-free version of ab32506.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y117
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-PKR antibody [Y117] (ab207341)
    • Alexa Fluor® 647 Anti-PKR antibody [Y117] (ab207342)
    • Anti-PKR antibody [Y117] (ab32506)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunohistochemistry kits

    • Rabbit specific HRP/DAB Detection IHC Detection Kit - Micro-polymer (ab236469)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HEK-293 whole cell lysate (ab7902)
  • Recombinant Protein

    • Recombinant human PKR protein (ab71666)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab239817 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

      ab32506 staining PKR in human liver carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

      Negative control 1: PBS in place of primary antibody.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32506).

    • Flow Cytometry (Intracellular) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Flow Cytometry (Intracellular) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

      Intracellular Flow Cytometry analysis of MCF-7 (human breast carcinoma) cells labeling PKR with purified ab32506 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32506). 

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

      ab32506 staining PKR in wild-type HAP1 cells (top panel) and PKR knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32506 at 1/400 dilution and ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.

      This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 4% formaldehyde (10 min).


      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32506).

    • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

      ab32506 staining PKR in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab32506 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

      Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
      Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32506).

    • Immunoprecipitation - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Immunoprecipitation - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

      ab32506 immunoprecipitating PKR. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

      Lane 1: HEK293 (human embryonic kidney) whole cell lysate (10ug)
      Lane 2: HEK293 (human embryonic kidney) whole cell lysate
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab32506 in HEK293 (human embryonic kidney) whole cell lysate

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32506).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

      Immunohistochemical analysis of paraffin-embedded human colon carcinoma using unpurified ab32506 at 1/100 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32506).

    • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

      Immunofluorescent staining of HeLa cells using unpurified ab32506 at 1/250 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32506).

    • Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)
      Anti-PKR antibody [Y117] - BSA and Azide free (ab239817)

    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 download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab239817? Please let us know so that we can cite the reference in this datasheet.

    ab239817 has not yet been referenced specifically in any publications.

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