Overview

  • Product name

    Anti-PERK antibody [EPR19876-294]
    See all PERK primary antibodies
  • Description

    Rabbit monoclonal [EPR19876-294] to PERK
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PERK aa 850 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9NZJ5

  • Positive control

    • WB: Wild-type HAP1, HeLa, U-87 MG, HEK-293T, MCF7, RAW 264.7, NIH/3T3 and PC-12 whole cell lysates; Mouse brain lysate; Rat brain and liver lysates.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab229912 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
WB 1/1000. Detects a band of approximately 150 kDa (predicted molecular weight: 125 kDa).
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.
  • Target

    • Function

      Phosphorylates the alpha subunit of eukaryotic translation-initiation factor 2 (EIF2), leading to its inactivation and thus to a rapid reduction of translational initiation and repression of global protein synthesis. Serves as a critical effector of unfolded protein response (UPR)-induced G1 growth arrest due to the loss of cyclin D1.
    • Tissue specificity

      Ubiquitous. A high level expression is seen in secretory tissues.
    • Involvement in disease

      Defects in EIF2AK3 are the cause of Wolcott-Rallison syndrome (WRS) [MIM:226980]; also known as multiple epiphyseal dysplasia with early-onset diabetes mellitus. WRS is a rare autosomal recessive disorder, characterized by permanent neonatal or early infancy insulin-dependent diabetes and, at a later age, epiphyseal dysplasia, osteoporosis, growth retardation and other multisystem manifestations, such as hepatic and renal dysfunctions, mental retardation and cardiovascular abnormalities.
    • Sequence similarities

      Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. GCN2 subfamily.
      Contains 1 protein kinase domain.
    • Domain

      The lumenal domain senses perturbations in protein folding in the ER, probably through reversible interaction with HSPA5/BIP.
    • Post-translational
      modifications

      Autophosphorylated.
      N-glycosylated.
    • Cellular localization

      Endoplasmic reticulum membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • DKFZp781H1925 antibody
      • E2AK3_HUMAN antibody
      • EC 2.7.11.1 antibody
      • Eif2ak3 antibody
      • Eukaryotic translation initiation factor 2 alpha kinase 3 antibody
      • Eukaryotic translation initiation factor 2-alpha kinase 3 antibody
      • Heme regulated EIF2 alpha kinase antibody
      • HRI antibody
      • HsPEK antibody
      • Pancreatic eIF2 alpha kinase antibody
      • Pancreatic eIF2-alpha kinase antibody
      • PEK antibody
      • PRKR like endoplasmic reticulum kinase antibody
      • PRKR-like endoplasmic reticulum kinase antibody
      • WRS antibody
      see all

    Images

    • All lanes : Anti-PERK antibody [EPR19876-294] (ab229912) at 1/1000 dilution

      Lane 1 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 2 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
      Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
      Lane 4 : Mouse brain lysate
      Lane 5 : Rat brain lysate
      Lane 6 : Rat liver lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 125 kDa
      Observed band size: 150 kDa
      why is the actual band size different from the predicted?



      Exposure time: Lanes 1-2: 37 seconds; Lanes 3-6: 3 minutes.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      This antibody also recognizes unidentifiable proteins below 37 kDa.

    • All lanes : Anti-PERK antibody [EPR19876-294] (ab229912) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : PERK knockout HAP1 whole cell lysate
      Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
      Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 5 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
      Lane 6 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 125 kDa
      Observed band size: 150 kDa why is the actual band size different from the predicted?


      Exposure time: 125 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      ab229912 was shown to specifically react with PERK in wild-type HAP1 cells as signal was lost in PERK knockout cells. Wild-type and PERK knockout samples were subjected to SDS-PAGE. ab229912 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

      This antibody also recognizes unidentifiable proteins below 75 kDa.

    References

    ab229912 has not yet been referenced specifically in any publications.

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