Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y116] to PKA R2/PKR2 - BSA and Azide free
  • Suitable for: IHC-P, WB, ICC, Flow Cyt (Intra), IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free
    See all PKA R2/PKR2 primary antibodies

  • Description

    Rabbit monoclonal [Y116] to PKA R2/PKR2 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    The antibody recognises PKA RII. It does not cross-react with other cAMP-dependent kinase. 

  • Tested applications

    Suitable for: IHC-P, WB, ICC, Flow Cyt (Intra), IPmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Cow

  • Immunogen

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

  • Positive control

    • WB: K562, HeLa and Jurkat cell lysates. IHC-P: Human colon cancer tissue. IP: K562 whole cell lysate. Flow Cyt (intra): MCF7 cells. ICC: HeLa cells.

  • General notes

    ab247265 is the carrier-free version of ab32514.

    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.

    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.

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

Properties

Images

  • Western blot - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    Western blot - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    All lanes : Anti-PKA R2/PKR2 antibody [Y116] (ab32514) at 1/10000 dilution (Purified)

    Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

    Predicted band size: 45 kDa



    This data was developed using ab32514, the same antibody clone in a different buffer formulation.

  • Immunoprecipitation - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    Immunoprecipitation - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)

    This data was developed using ab32514, the same antibody clone in a different buffer formulation.
    Purified ab32514 at 1/20 dilution (0.5µg) immunoprecipitating PKA R2/PKR2 in K-562 whole cell lysate.
    Lane 1 (input): K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
    Lane 2 (+): ab32514 + K-562 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32514 in K-562 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: 50 kDa

  • Flow Cytometry - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    Flow Cytometry - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    This data was developed using ab32514, the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling PKA R2/PKR2 with purified ab32514 at 1/20 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).
  • Immunocytochemistry - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    Immunocytochemistry - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    This data was developed using ab32514, the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKA R2/PKR2 with purified ab32514 at 1/500 dilution (1.9 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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    This data was developed using ab32514, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon cancer tissue sections labeling PKA R2/PKR2 with purified ab32514 at 1/100 dilution (0.84 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.
  • Western blot - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    Western blot - Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    All lanes : Anti-PKA R2/PKR2 antibody [Y116] (ab32514) at 1/1000 dilution (Unpurified)

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PRKAR2A knockout HeLa 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: 45 kDa
    Observed band size: 50 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 (ab32514).

    Lanes 1-3: Merged signal (red and green). Green - ab32514 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa. 

    Unpurified ab32514 Anti-PKA R2/PKR2 antibody [Y116] was shown to specifically react with PKA R2/PKR2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265748 (knockout cell lysate ab257607) was used.  Wild-type and PKA R2/PKR2 knockout samples were subjected to SDS-PAGE.  ab32514 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.

  • Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)
    Anti-PKA R2/PKR2 antibody [Y116] - BSA and Azide free (ab247265)

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