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Validated using a knockout cell lineRecombinant

Recombinant Anti-PKC alpha antibody [133] (ab11723)

Reviews (1) Submit a question References (17)
Western blot - Anti-PKC alpha antibody [133] (ab11723)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)
  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)
  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [133] to PKC alpha
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-PKC alpha antibody [133]
    See all PKC alpha primary antibodies

  • Description

    Mouse monoclonal [133] to PKC alpha

  • Host species

    Mouse

  • Specificity

    ab11723 recognises the alpha isoform of PKC.

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab55672)

  • Positive control

    • WB: HAP1, K562, C6, HeLa, NIH/3T3 and HEK-293 cell lysate. Human kidney tissue lysate. Mouse and rat brain and spleen tissue lysate. Recombinant human PKC alpha protein (Active) (ab55672). IHC-P: Human, mouse and rat retina tissue.

  • General notes

    This product has switched from a hybridoma to recombinant production method on 25th July 2021.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab11723 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (1)
1/500 - 1/1000. Predicted molecular weight: 76 kDa.Can be blocked with Recombinant human PKC alpha protein (Active) (ab55672).
IHC-P
1/1 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/500 - 1/1000. Predicted molecular weight: 76 kDa.Can be blocked with Recombinant human PKC alpha protein (Active) (ab55672).
IHC-P
1/1 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC.

Target

  • Function

    This is a calcium-activated, phospholipid-dependent, serine- and threonine-specific enzyme. May play a role in cell motility by phosphorylating CSPG4.
    PKC is activated by diacylglycerol which in turn phosphorylates a range of cellular proteins. PKC also serves as the receptor for phorbol esters, a class of tumor promoters.

  • Sequence similarities

    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 C2 domain.
    Contains 2 phorbol-ester/DAG-type zinc fingers.
    Contains 1 protein kinase domain.

  • Cellular localization

    Cytoplasm. Cell membrane. Nucleus.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • AAG6 antibody
    • Aging associated gene 6 antibody
    • aPKC antibody
    • KPCA_HUMAN antibody
    • PKC alpha antibody
    • PKC-A antibody
    • PKC-alpha antibody
    • PKCA antibody
    • PRKACA antibody
    • PRKCA antibody
    • Protein Kinase C alpha antibody
    • Protein kinase C alpha type antibody
    see all

Images

  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
    Western blot - Anti-PKC alpha antibody [133] (ab11723)

    This image was generated from the hybridoma version of the product.

    Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 6 and 10: PKC alpha knockout HAP1 cell lysate (20 µg)
    Lanes 3, 7 and 11: K562 cell lysate (20 µg)
    Lanes 4, 8 and 12: HEK293 cell lysate (20 µg)
    Lanes 1, 2, 3 and 4: Green signal from target – ab11723 observed at 77 kDa
    Lanes 5, 6, 7 and 8: Red signal from loading control – ab8245 observed at 37 kDa
    Lanes 9, 10, 11 and 12: Merged (red and green) signal

    ab11723 was shown to specifically react with PKC alpha in wild-type HAP1 cells. No band was observed when PKC alpha knockout samples were examined. Wild-type and PKC alpha knockout samples were subjected to SDS-PAGE. ab11723 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)

    Immunohistochemical analysis of Paraffin-embedded human retina tissue labelling PKC alpha with ab11723 at 1/500 dilution, followed by ready to use secondary antibody Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879). Positive staining on human retina is observed. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Goat Anti-Mouse IgG H&L (HRP polymer) ab214879.

     

  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
    Western blot - Anti-PKC alpha antibody [133] (ab11723)

    This image was generated from the hybridoma version of the product.

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PKC alpha knockout HAP1 cell lysate (20 µg)
    Lane 3: K562 cell lysate (20 µg)
    Lane 4: HEK293 cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab11723 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab11723 and a competitor's top cited rabbit polyclonal antibody.

  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
    Western blot - Anti-PKC alpha antibody [133] (ab11723)
    All lanes : Anti-PKC alpha antibody [133] (ab11723) at 1/1000 dilution

    Lane 1 : Recombinant human PKC alpha protein (Active) (ab55672)
    Lane 2 : Recombinant human PKC beta 1 protein (ab60840)
    Lane 3 : Recombinant human PKC beta 2 protein (ab60841)
    Lane 4 : Recombinant human PKC eta protein (ab60849)
    Lane 5 : Recombinant human PKC epsilon protein (ab60847)
    Lane 6 : Recombinant human PKC theta/PRKCQ protein (ab56641)
    Lane 7 : Recombinant human PKC mu/PKD protein (ab60873)
    Lane 8 : Recombinant human PKC zeta protein (ab60848)
    Lane 9 : Recombinant human PKC iota protein (ab60850)
    Lane 10 : Recombinant human PKC gamma protein (ab60842)

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

    Predicted band size: 76 kDa



    5% NFDM/TBST

    All positive controls 10 ng

  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
    Western blot - Anti-PKC alpha antibody [133] (ab11723)
    All lanes : Anti-PKC alpha antibody [133] (ab11723) at 1/1000 dilution

    Lane 1 : Recombinant human PKC alpha protein (Active) (ab55672)
    Lane 2 : Recombinant human PKC delta protein (ab60844)

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

    Predicted band size: 76 kDa


    Exposure time: 180 seconds


    5% NFDM/TBST

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)

    Immunohistochemical analysis of paraffin-embedded mouse retina tissue labelling PKC alpha with ab11723 at 1/500 dilution, followed by ready to use secondary antibody Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879). Positive staining on mouse retina is observed. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Goat Anti-Mouse IgG H&L (HRP polymer) ab214879.

  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
    Western blot - Anti-PKC alpha antibody [133] (ab11723)
    All lanes : Anti-PKC alpha antibody [133] (ab11723) at 1/1000 dilution

    Lane 1 : Human kidney tissue lysate 20 ug
    Lane 2 : Mouse brain tissue lysate 20 ug
    Lane 3 : Mouse spleen tissue lysate 20 ug
    Lane 4 : Rat brain tissue lysate 20 ug
    Lane 5 : Rat spleen tissue lysate 20 ug

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

    Predicted band size: 76 kDa


    Exposure time: 180 seconds


    5% NFDM/TBST

  • Western blot - Anti-PKC alpha antibody [133] (ab11723)
    Western blot - Anti-PKC alpha antibody [133] (ab11723)
    All lanes : Anti-PKC alpha antibody [133] (ab11723) at 1/1000 dilution

    Lane 1 : C6 (rat glial tumor glial cell), whole cell lysate 20 µg
    Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell ), whole cell lysate 20 µg
    Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 20 µg

    Secondary
    All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

    Predicted band size: 76 kDa



    Exposure time

    Lane 1: 136 seconds
    Lane 2 and 3: 15 seconds

    Fresh lysates were used in lane 2 and 3.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC alpha antibody [133] (ab11723)

    Immunohistochemical analysis of paraffin-embedded rat retina tissue labelling PKC alpha with ab11723 at 1/500 dilution, followed by ready to use secondary antibody Goat Anti-Mouse IgG H&L (HRP polymer) (ab214879). Positive staining on rat retina is observed. Counter stained with Hematoxylin. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0)

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Goat Anti-Mouse IgG H&L (HRP polymer) ab214879.

References (17)

Publishing research using ab11723? Please let us know so that we can cite the reference in this datasheet.

ab11723 has been referenced in 17 publications.

  • Hu S  et al. In Vivo CRISPR/Cas9-Mediated Genome Editing Mitigates Photoreceptor Degeneration in a Mouse Model of X-Linked Retinitis Pigmentosa. Invest Ophthalmol Vis Sci 61:31 (2020). PubMed: 32330228
  • Guo Y  et al. Modeling Retinitis Pigmentosa: Retinal Organoids Generated From the iPSCs of a Patient With the USH2A Mutation Show Early Developmental Abnormalities. Front Cell Neurosci 13:361 (2019). PubMed: 31481876
  • Orlandi C  et al. Transsynaptic Binding of Orphan Receptor GPR179 to Dystroglycan-Pikachurin Complex Is Essential for the Synaptic Organization of Photoreceptors. Cell Rep 25:130-145.e5 (2018). PubMed: 30282023
  • De Silva SR  et al. Long-term restoration of visual function in end-stage retinal degeneration using subretinal human melanopsin gene therapy. Proc Natl Acad Sci U S A 114:11211-11216 (2017). IHC-P . PubMed: 28973921
  • Wang Y  et al. The Auxiliary Calcium Channel Subunit a2d4 Is Required for Axonal Elaboration, Synaptic Transmission, and Wiring of Rod Photoreceptors. Neuron 93:1359-1374.e6 (2017). PubMed: 28262416
View all Publications for this product

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Western blot abreview for Anti-PKC alpha antibody [133]

 Good
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cells)
Specification
HeLa cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Dec 05 2005

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