Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa, 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

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 HeLa cell lysate
    Lane 2 : PRKRA knockout HeLa 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 HeLa 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)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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