Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2730Y] to PKC (phospho T514) - BSA and Azide free
  • Suitable for: IHC-P, IP, WB
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free
    See all PKC primary antibodies

  • Description

    Rabbit monoclonal [EP2730Y] to PKC (phospho T514) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • General notes

    ab247898 is the carrier-free version of ab109539.

    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-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) at 1/2000 dilution (purified)

    Lane 1 : Rat brain lysates
    Lane 2 : Rat brain lysates and then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 78 kDa



    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Immunoprecipitation - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)

    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    ab109539 (purified) at 1:20 dilution (2µg) immunoprecipitating PKC in Rat brain lysate.
    Lane 1 (input): Rat brain lysate 10µg
    Lane 2 (+): ab109539 & Rat brain lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109539 in Rat brain lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)

    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebellum tissue sections labeling PKC with Purified ab109539 at 1:300 dilution (1.14 µg/ml). Heat mediated antigen retrieval was performed using citrate buffer, pH 6.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)

    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebellum tissue sections labeling PKC with Purified ab109539 at 1:300 dilution (1.14 µg/ml). Heat mediated antigen retrieval was performed using citrate buffer, pH 6.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)

    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebral cortex tissue sections labeling PKC with Purified ab109539 at 1:300 dilution (1.14 µg/ml). Heat mediated antigen retrieval was performed using citrate buffer, pH 6.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
  • Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) at 1/2000 dilution (purified)

    Lane 1 : Mouse cerebellum lysates
    Lane 2 : Mouse cerebellum lysates and then the membrane was incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 78 kDa



    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) at 1/2000 dilution (unpurified)

    Lane 1 : Active human PKC alpha full length protein.
    Lane 2 : Active human PKC beta 1 full length protein.
    Lane 3 : Active human PKC beta 2 full length protein.
    Lane 4 : Active human PKC delta full length protein.
    Lane 5 : Active human PKC eta full length protein.
    Lane 6 : Active human PKC epsilon full length protein.
    Lane 7 : Active human PKC theta full length protein.
    Lane 8 : Active human PKC mu full length protein.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 78 kDa
    Observed band size: 100-150 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    Active human PKC alpha full length protein (ab55672) contains aa1-672 with GST-tag; Active human PKC beta 1 full length protein (ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein (ab60841) contains aa1-673 with GST-tag; Active human PKC delta full length protein (ab60844) contains aa1-676 with GST-tag; Active human PKC eta full length protein (ab60849) contains aa1-683 with GST-tag; Active human PKC epsilon full length protein (ab60847) contains aa1-737 with GST-tag; Active human PKC theta full length protein (ab56641) contains aa1-706 with GST-tag; Active human PKC mu full length protein (ab60873) contains aa1-912 with GST-tag.

  • Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) at 1/5000 dilution (purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Phorbol-12-myristate-13-acetate whole cell lysates
    Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Phorbol-12-myristate-13-acetate whole cell lysates. The membrane was then incubated with phosphatase.

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 78 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Western blot - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) at 0.02 µg/ml (unpurified)

    All lanes : HeLa cell lysate

    Predicted band size: 78 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    This data was developed using ab109539, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    This data was developed using ab109539, the same antibody clone in a different buffer formulation.Unpurified ab109539 at 1/100 dilution staining PKC (phospho T514) in paraffin-embedded Human brain tissue by Immunohistochemistry. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)
    Anti-PKC (phospho T514) antibody [EP2730Y] - BSA and Azide free (ab247898)

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