Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)

Key features and details

  • Rabbit monoclonal [EP2608Y] to PKC alpha (phospho T497)
  • Suitable for: Dot blot, WB, IHC-P
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-PKC alpha (phospho T497) antibody [EP2608Y]
    See all PKC alpha primary antibodies

  • Description

    Rabbit monoclonal [EP2608Y] to PKC alpha (phospho T497)

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

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

  • Immunogen

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

  • Positive control

    • WB: Jurkat cell lysate, HeLa human cervix adenocarcinoma cells treated with Calyculin A whole cell lysates and K562 human chronic myelogenous leukemia cells treated with Calyculin A whole cell lysates. IHC-P: Human kidney tissue.

  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

Images

  • Western blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    Western blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    All lanes : Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016) at 1/20000 dilution

    Lane 1 : Untreated K562 (human chronic myelogenous leukemia) whole cell lysates
    Lane 2 : K562 (human chronic myelogenous leukemia) cells treated with Calyculin A whole cell lysates
    Lane 3 : K562 (human chronic myelogenous leukemia) cells were treated with Calyculin A whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa


    Exposure time: 3 minutes


    Blocking and diluting buffer - 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)

    ab76016, at 1/100 dilution, staining PKC alpha in human kidney by immunohistochemistry using paraffin-embedded tissue.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    Western blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    All lanes : Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016) at 1/20000 dilution

    Lane 1 : Untreated HeLa (human cervix adenocarcinoma) whole cell lysates
    Lane 2 : HeLa (human cervix adenocarcinoma) cells treated with Calyculin A whole cell lysates
    Lane 3 : HeLa (human cervix adenocarcinoma) cells were treated with Calyculin A whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa


    Exposure time: 3 minutes


    Blocking/Diluting Buffer - 5% NFDM/TBST

  • Western blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    Western blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    All lanes : Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016) at 1/100000 dilution

    Lane 1 : Jurkat cell lysate - untreated
    Lane 2 : Jurkat cell lysate - treated with LP

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/1000 dilution

    Predicted band size: 77 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?



    LP is Lambda Phosphatase. It is not an inducer of phosphorylation but is essential to treat the membrane (not the cells).

  • Dot Blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)
    Dot Blot - Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)

    Dot blot analysis of PKC alpha (pT497) peptide (Lane 1) and PKC alpha non-phospho peptide (Lane 2) labelling PKC alpha (phospho T497) with ab76016 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

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