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Validated using a knockout cell lineRecombinantRabMAb

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

Submit a review Submit a question References (11)
Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
  • Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
  • Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
  • Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3224] to PACT (PKR activating protein) / PRKRA
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide within Human PACT (PKR activating protein)/ PRKRA aa 300-400 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HEK-293T, K562, Jurkat and HepG2 whole cell lysate (ab7900). IHC-P: Human brain tissue.

  • General notes

    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.

Properties

Applications

The Abpromise guarantee

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
Notes
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

    see all

  • 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

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

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : PRKRA knockout HEK-293T cell lysate
    Lane 3 : K-562 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 34 kDa
    Observed band size: 36 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab75749 observed at 36 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab75749 Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] was shown to specifically react with PACT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266806 (knockout cell lysate ab258141) was used.  Wild-type and PACT knockout samples were subjected to SDS-PAGE.  ab75749 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)

    ab75749 at 1/100 dilution staining PACT (PKR activating protein) / PRKRA in paraffin-embedded human brain tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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

    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.

  • Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
    Western blot - Anti-PACT (PKR activating protein) / PRKRA antibody [EPR3224] (ab75749)
    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
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 34 kDa
    Observed band size: 34 kDa

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

References (11)

Publishing research using ab75749? Please let us know so that we can cite the reference in this datasheet.

ab75749 has been referenced in 11 publications.

  • Ye T  et al. Large extracellular vesicles secreted by human iPSC-derived MSCs ameliorate tendinopathy via regulating macrophage heterogeneity. Bioact Mater 21:194-208 (2023). PubMed: 36101856
  • Vaughn LS  et al. DYT-PRKRA Mutation P222L Enhances PACT's Stimulatory Activity on Type I Interferon Induction. Biomolecules 12:N/A (2022). PubMed: 35625640
  • Montavon TC  et al. Human DICER helicase domain recruits PKR and modulates its antiviral activity. PLoS Pathog 17:e1009549 (2021). PubMed: 33984068
  • Ukhueduan B  et al. Regulation of PKR activation and apoptosis during oxidative stress by TRBP phosphorylation. Int J Biochem Cell Biol 137:106030 (2021). PubMed: 34174402
  • Suzuki S  et al. Dental pulp cell-derived powerful inducer of TNF-a comprises PKR containing stress granule rich microvesicles. Sci Rep 9:3825 (2019). PubMed: 30846715
View all Publications for this product

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