Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [YE350] to PKR - BSA and Azide free
  • Suitable for: ICC, IP, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PKR antibody [YE350] - BSA and Azide free
    See all PKR primary antibodies

  • Description

    Rabbit monoclonal [YE350] to PKR - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, IP, IHC-P, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A549, K562, HAP1, HepG2, Jurkat, and HeLa cell lysates. IP: Jurkat cell lysate IHC: Human cerebrum tissue; ICC: MCF7 cells; Flow Cyt: HeLa cells.

  • General notes

    ab239799 is the carrier-free version of ab32052 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    Ab239799 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

    Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : EIF2AK2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 4 : EIF2AK2 knockout A549 (Human lung carcinoma cell line) whole cell lysate
    Lane 5 : K562 (Human chronic myelogenous leukemia lymphoblast cell line) whole 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: 62 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab32052, the same antibody clone in a different buffer formulation.

    Lanes 1-5: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261824 (knockout cell lysate ab256899) was used. Wild-type and PKR knockout samples were subjected to SDS-PAGE. ab32052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 paraffin embedded sections - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Immunohistochemistry paraffin embedded sections - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    This data was developed using ab32052 the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling PKR with Purified ab32052 at 1:100 dilution (2.52 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Immunocytochemistry - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Immunocytochemistry - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)

    This data was developed using ab32052 the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PKR with Purified ab32052 at 1:50 dilution (5.04 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Flow Cytometry - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    This data was developed using ab32052 the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKR with Purified ab32052 at 1:30 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Immunoprecipitation - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Immunoprecipitation - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)

    This data was developed using ab32052, the same antibody clone in a different buffer formulation.
    Purified ab32052 at 1/20 dilution (1µg) immunoprecipitating PKR in Jurkat whole cell lysate.
    Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10µg
    Lane 2 (+): ab32052 + Jurkat whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32052 in Jurkat whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 68 kDa
    Possible degradation bands are observed between 20-30kDa.

  • Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Anti-PKR antibody [YE350] (ab32052) at 1/5000 dilution + Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 62 kDa



    This data was developed using ab32052, the same antibody clone in a different buffer formulation.

  • Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : EIF2AK2 knockout A549 cell lysate
    Lane 3 : K-562 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: 62 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab32052).

    Lanes 1-3: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267000 (knockout cell lysate ab256901) was used.  Wild-type and PKR knockout samples were subjected to SDS-PAGE.  ab32052 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.

  • Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    Western blot - Anti-PKR antibody [YE350] - BSA and Azide free (ab239799)
    All lanes : Anti-PKR antibody [YE350] (ab32052) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : EIF2AK2 knockout A549 cell lysate
    Lane 3 : K-562 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: 62 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab32052).

    Lanes 1-3: Merged signal (red and green). Green - ab32052 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab32052 Anti-PKR antibody [YE350] was shown to specifically react with PKR in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266999 (knockout cell lysate ab256900) was used.  Wild-type and PKR knockout samples were subjected to SDS-PAGE.  ab32052 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.

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