製品の概要

  • 製品名
    Anti-PKR antibody [YE350]
    PKR 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [YE350] to PKR
  • 特異性
    ab32052 recognises Protein kinase R (PKR). It does not cross-react with other GCN2 family members.
  • アプリケーション
    適用あり: Flow Cyt, WB, IHC-P, ICC/IF, IPmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PKR aa 50-150.

  • ポジティブ・コントロール
    • WB: HeLa and MCF-7 cell lysates. ICC/IF: HeLa cells. IHC-P: Human stomach carcinoma and human liver tissues. Flow Cyt: MCF-7 cells.
  • 特記事項

    Mouse, 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.

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • バッファー
    PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • 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
    YE350
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab32052 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Flow Cyt 1/20.
WB 1/1000 - 1/10000. Detects a band of approximately 68 kDa (predicted molecular weight: 62 kDa).
IHC-P 1/100.
ICC/IF 1/100 - 1/500.
IP 1/20 - 1/100.

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

Anti-PKR antibody [YE350] 画像



  • Predicted band size : 62 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: EIF2AK2 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: K652 whole cell lysate (20 µg)
    Lane 4: HepG2 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab32052 was shown to specifically react with EIF2AK2 when EIF2AK2 knockout samples were used. Wild-type and EIF2AK2 knockout samples were subjected to SDS-PAGE. Ab32052 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Anti-PKR antibody [YE350] (ab32052) at 1/10000 dilution (purified) + MCF-7 whole cell lysate at 10 µg

    Secondary
    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?)
  • Immunofluorescence staining of HeLa cells with purified ab32052 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32052 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human liver with purified ab32052 at a working dilution of 1/100. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Flow Cytometry analysis of MCF-7 cells labelling PKR with purified ab32052 at a dilution of 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Flour® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Anti-PKR antibody [YE350] (ab32052) at 1/10000 dilution (purified) + HeLa whole cell lysate at 10 µg

    Secondary
    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?)
  • Immunohistochemical analysis of paraffin-embedded human stomach carcinoma using unpurified ab32052 at 1/250 dilution.

Anti-PKR antibody [YE350] (ab32052) 使用論文

This product has been referenced in:
  • Hu Z  et al. Silencing nc886, a Non-Coding RNA, Induces Apoptosis of Human Endometrial Cancer Cells-1A In Vitro. Med Sci Monit 23:1317-1324 (2017). Read more (PubMed: 28298621) »
  • Chang YH  et al. dsRNA Binding Domain of PKR Is Proteolytically Released by Enterovirus A71 to Facilitate Viral Replication. Front Cell Infect Microbiol 7:284 (2017). Read more (PubMed: 28702377) »

See all 14 Publications for this product

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LOT:GR56038-1


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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Mirjam Eckert

Verified customer

投稿 Mar 26 2009

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

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