Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16794] to PKC - BSA and Azide free
  • Suitable for: ICC, WB, IP, IHC-P
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

Overview

  • Product name

    Anti-PKC antibody [EPR16794] - BSA and Azide free
    See all PKC primary antibodies

  • Description

    Rabbit monoclonal [EPR16794] to PKC - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human, Recombinant fragment

  • Immunogen

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

  • General notes

    ab250107 is the carrier-free version of ab179521 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

    Ab250107 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Recombinant PKC isoforms were tested for reactivity to ab179521 by western blot, and are marked as positive (+) or negative (-).

    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.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Western blot - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    All lanes : Anti-PKC antibody [EPR16794] (ab179521) at 1/2000 dilution

    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 gamma full length protein
    Lane 5 : Active human PKC delta full length protein
    Lane 6 : Active human PKC eta full length protein
    Lane 7 : Active human PKC epsilon full length protein
    Lane 8 : Active human PKC theta full length protein
    Lane 9 : Active human PKC mu full length protein

    Lysates/proteins at 0.02 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 77, 78 kDa


    Exposure time: 5 seconds


    This data was developed using ab179521, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    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 gamma full length protein (ab60842) contains aa1-677 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 antibody [EPR16794] - BSA and Azide free (ab250107)
    Western blot - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Anti-PKC antibody [EPR16794] (ab179521) at 1/2000 dilution + Active human PKC gamma full length protein at 0.02 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 77, 78 kDa


    Exposure time: 3 minutes


    This data was developed using ab179521, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Active human PKC gamma full length protein (ab60842) contains aa1-677 with GST-tag.

    ab179521 weakly recognizes PKC gamma, and is only evident at long exposure times.

  • Other - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Other - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)

    This data was developed using ab179521, the same antibody clone in a different buffer formulation.

    Recombinant PKC isoforms were tested for reactivity by western blot, and are marked as positive (+) or negative (-).
  • Western blot - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Western blot - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    All lanes : Anti-PKC antibody [EPR16794] (ab179521) at 1/5000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal kidney lysate
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 77, 78 kDa
    Observed band size: 78 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab179521, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Western blot - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Anti-PKC antibody [EPR16794] (ab179521) at 1/5000 dilution + L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 77, 78 kDa
    Observed band size: 78 kDa why is the actual band size different from the predicted?



    This data was developed using ab179521, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Immunocytochemistry - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    This data was developed using ab179521, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling PKC alpha+beta1+beta2 with ab179521 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear and cytoplasm staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).The negative controls are as follows:
    -ve control 1: ab179521 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    This data was developed using ab179521, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Human breast cancer is observed. Counter stained with Hematoxylin. Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    This data was developed using ab179521, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse liver is observed. Counter stained with Hematoxylin. Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    This data was developed using ab179521, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PKC alpha+beta1+beta2 with ab179521 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on rat testis is observed. Counter stained with Hematoxylin. Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Immunoprecipitation - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Immunoprecipitation - Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    This data was developed using ab179521, the same antibody clone in a different buffer formulation.PKC alpha+beta1+beta2 was immunoprecipitated from 1mg of Mouse brain whole cell lysate with ab179521 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab179521 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
    Lane 1: Mouse brain whole cell lysate 10µg (Input).
    Lane 2: ab179521 IP in Mouse brain whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab179521 in Mouse brain whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 1 second.
  • Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)
    Anti-PKC antibody [EPR16794] - BSA and Azide free (ab250107)

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