Validated using a knockout cell line
Recombinant
RabMAb

Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)

Overview

  • Product name
    Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224]
    See all PACT (PKR activating protein) / PRKRA primary antibodies
  • Description
    Rabbit monoclonal [EPR3224] to PACT (PKR activating protein) / PRKRA
  • Tested applications
    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues from the C terminal of human PACT (PKR activating protein)/ PRKRA

  • Positive control
    • K562, Jurkat and HepG2 cell lysates. Human brain tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab75749 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
IHC-P
  • Application notes
    IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval using 0.01M Sodium Citrate Buffer, pH 6.0 before commencing with IHC staining protocol.
    WB: 1/1000 - 1/5000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).

    Is unsuitable for Flow Cyt, ICC or IP.


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Activates EIF2AK2/PKR in the absence of double stranded RNA (dsRNA), leading to phosphorylation of EIF2S1/EFI2-alpha and inhibition of translation and induction of apoptosis. Required for siRNA production by DICER1 and for subsequent siRNA-mediated post-transcriptional gene silencing. Does not seem to be required for processing of pre-miRNA to miRNA by DICER1.
    • Involvement in disease
      Defects in PRKRA are the cause of dystonia type 16 (DYT16) [MIM:612067]. DYT16 is an early-onset dystonia-parkinsonism disorder. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYT16 patients have progressive, generalized dystonia with axial muscle involvement, oro-mandibular (sardonic smile) and laryngeal dystonia and, in some cases, parkinsonian features.
    • Sequence similarities
      Belongs to the PRKRA family.
      Contains 3 DRBM (double-stranded RNA-binding) domains.
    • Domain
      Self-association may occur via interactions between DRBM domains as follows: DRBM 1/DRBM 1, DRBM 1/DRBM 2, DRBM 2/DRBM 2 or DRBM 3/DRBM3.
    • Post-translational
      modifications
      Phosphorylated at Ser-246 in unstressed cells and at Ser-287 in stressed cells. Phosphorylation at Ser-246 appears to be a prerequisite for subsequent phosphorylation at Ser-287. Phosphorylation at Ser-246 and Ser-287 are necessary for activation of EIF2AK2/PKR under conditions of stress.
    • Cellular localization
      Cytoplasm > perinuclear region.
    • Information by UniProt
    • Database links
    • Alternative names
      • DYT16 antibody
      • HSD14 antibody
      • Interferon inducible double stranded RNA dependent protein kinase activator A antibody
      • interferon-inducible double stranded RNA-dependent activator antibody
      • Interferon-inducible double stranded RNA-dependent protein kinase activator A antibody
      • PACT antibody
      • PKR associated protein X antibody
      • PKR associating protein X antibody
      • PKR-associated protein X antibody
      • PKR-associating protein X antibody
      • PRKRA antibody
      • PRKRA_HUMAN antibody
      • Protein activator of the interferon induced protein kinase antibody
      • Protein activator of the interferon-induced protein kinase antibody
      • Protein kinase antibody
      • Protein kinase interferon inducible double stranded RNA dependent activator antibody
      • RAX antibody
      see all

    Images



    • Predicted band size : 34 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: PACT (PKR activating protein)/PRKRA knockout HAP1 cell lysate (20 µg)
      Lane 3: K562 cell lysate (20 µg)
      Lane 4: HepG2 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab75749 observed at 36 kDa. Red - loading control, ab18058, observed at 124 kDa.

      ab75749 was shown to specifically react with PACT (PKR activating protein)/PRKRA when PACT (PKR activating protein)/PRKRA knockout samples were used. Wild-type and PACT (PKR activating protein)/PRKRA knockout samples were subjected to SDS-PAGE. ab75749 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749) at 1/1000 dilution

      Lane 1 : K562 cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : HepG2 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 34 kDa
      Observed band size : 34 kDa
    • ab75749 at 1/100 dilution staining PACT (PKR activating protein) / PRKRA in paraffin-embedded human brain tissue.

    References

    This product has been referenced in:
    • Emde A  et al. Dysregulated miRNA biogenesis downstream of cellular stress and ALS-causing mutations: a new mechanism for ALS. EMBO J 34:2633-51 (2015). Read more (PubMed: 26330466) »
    • Gross TJ  et al. A microRNA processing defect in smokers' macrophages is linked to SUMOylation of the endonuclease DICER. J Biol Chem 289:12823-34 (2014). WB ; Human . Read more (PubMed: 24668803) »

    See all 3 Publications for this product

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