Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19374] to PKR - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PKR antibody [EPR19374] - BSA and Azide free
    See all PKR primary antibodies

  • Description

    Rabbit monoclonal [EPR19374] to PKR - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A459, K562, Jurkat, HeLa, MCF7, 4T1, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, cerebral cortex, hippocampus, lung, thymus and heart lysates; Rat brain, cerebral cortex, heart and spleen lysates. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.

  • General notes

    ab224887 is the carrier-free version of ab184257.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    Western blot - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    All lanes : Anti-PKR antibody [EPR19374] (ab184257) 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: 58 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab184257, the same antibody clone in a different buffer formulation.

    Lanes 1-5: Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab184257 Anti-PKR antibody [EPR19374] 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. ab184257 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.

  • Flow Cytometry - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    Flow Cytometry - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)

    Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PKR with ab184257 at 1/120 dilution (red) compared with a Rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti Rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184257).

  • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKR with ab184257 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasm and weakly nuclear staining on HeLa cell line. 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [EPR19374] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab184257 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184257).

  • Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    Immunocytochemistry/ Immunofluorescence - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 Mouse embryonic fibroblast cell line) cells labeling PKR with ab184257 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasm and weakly nuclear staining on NIH/3T3 cell line. 

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [EPR19374]- Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab184257 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184257).

  • Western blot - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    Western blot - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    All lanes : Anti-PKR antibody [EPR19374] (ab184257) 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: 58 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 (ab184257).

    Lanes 1-3: Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab184257 Anti-PKR antibody [EPR19374] 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.  ab184257 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 [EPR19374] - BSA and Azide free (ab224887)
    Western blot - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    All lanes : Anti-PKR antibody [EPR19374] (ab184257) 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: 58 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 (ab184257).

    Lanes 1-3: Merged signal (red and green). Green - ab184257 observed at 70 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab184257 Anti-PKR antibody [EPR19374] 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.  ab184257 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.

  • Immunoprecipitation - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    Immunoprecipitation - Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)

    PKR was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab184257 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab184257 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10µg (Input).

    Lane 2: ab184257 IP in NIH/3T3 whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR19374] - Isotype Control (ab172730) instead of ab184257 in NIH/3T3 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184257).

  • Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)
    Anti-PKR antibody [EPR19374] - BSA and Azide free (ab224887)

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