Anti-PKC delta (phospho S645) antibody [EP1486Y] (ab108972)

Overview

  • Product name
    Anti-PKC delta (phospho S645) antibody [EP1486Y]
    See all PKC delta primary antibodies
  • Description
    Rabbit monoclonal [EP1486Y] to PKC delta (phospho S645)
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human PKC delta aa 600 to the C-terminus (phospho S645).
    Database link: Q05655

  • Positive control
    • 293T cell lysate; Human breat carcinoma; HeLa cells
  • General notes

    A trial size is available to purchase for this antibody.

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

    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

Associated products

Applications

Our Abpromise guarantee covers the use of ab108972 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 1/10000 - 1/50000. Predicted molecular weight: 78 kDa.
IHC-P 1/50 - 1/100. Antigen retrieval is recommended.
ICC/IF 1/50 - 1/100.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function
      This is calcium-independent, phospholipid-dependent, serine- and threonine-specific enzyme. 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. May play a role in antigen-dependent control of B-cell function. Phosphorylates MUC1 in the C-terminal and regulates the interaction between MUC1 and beta-catenin.
    • 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.
    • Domain
      The C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor.
      The C2 domain is a non-calcium binding domain. It binds proteins containing phosphotyrosine in a sequence-specific manner.
    • Post-translational
      modifications
      Phosphorylated on Thr-507, within the activation loop. Autophosphorylated and/or phosphorylated. Although the Thr-507 phosphorylation occurs it is not a prerequisite for enzymatic activity.
    • Cellular localization
      Cytoplasm. Membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • CVID9 antibody
      • D14Ertd420e antibody
      • Kinase PKC delta antibody
      • KPCD antibody
      • KPCD_HUMAN antibody
      • MAY 1 antibody
      • MAY1 antibody
      • MGC49908 antibody
      • nPKC delta antibody
      • nPKC-delta antibody
      • PCKd antibody
      • PKC d antibody
      • PKC delta antibody
      • PKCD antibody
      • PKCdelta antibody
      • PRKC D antibody
      • PRKC delta antibody
      • Prkcd antibody
      • Protein Kinase C delta antibody
      • Protein kinase C delta type antibody
      • Protein kinase C delta VIII antibody
      • Protein Kinase Cdelta antibody
      • Tyrosine protein kinase PRKCD antibody
      see all

    Images



    • Predicted band size : 78 kDa

      Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 6 and 10: PKC delta knockout HAP1 cell lysate (20 µg)
      Lanes 3, 7 and 11: A431 cell lysate (20 µg)
      Lanes 4, 8 and 12: HeLa cell lysate (20 µg)
      Lanes 1, 2, 3 and 4: Green signal from target – ab108972 observed at 78 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

      ab108972 was shown to specifically react with PKC delta when PKC delta knockout samples were used. Wild-type and PKC delta knockout samples were subjected to SDS-PAGE. ab108972 and ab8245 (loading control to GAPDH) were diluted 1/10000 and 1/2000 respectively and incubated overnight at 4ºC. 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 1/10000 dilution for 1 h at room temperature before imaging.

    • Immunofluorescent staining of PKC delta in HeLa cells, using ab108972 at a 1/50 dilution.
    • Immunohistochemical analysis of PKC delta in paraffin embedded Human breast carcinoma tissue, using ab108972 at a 1/50 dilution.


    • Predicted band size : 78 kDa

      Primary ab Dilution 1:100,000 dilution, Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution 1:20,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM/TBST, Lane 1: THP-1 whole cell lysate treated with Calyculin A with no peptides, Lane 2: THP-1 whole cell lysate treated with Calyculin A with PKCd S645Pho peptides, Lane 3: THP-1 whole cell lysate treated with Calyculin A with PKCd unmodified peptides, Lane 4: None, Observed MW   78 kDa, Exposure time   10 seconds



    • Predicted band size : 78 kDa

      Primary ab Dilution 1:100,000 dilution, Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution 1: 20,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM/TBST, Lane 1: Untreated THP-1 (Human monocytic leukemia cell line) whole cell lysates 15ug, Lane 2: THP-1 (Human monocytic leukemia cell line) treated with 50nM Calyculin A for 60 minutes whole cell lysates 15ug. Lane 3: THP-1 (Human monocytic leukemia cell line) treated with 50nM Calyculin A for 60 minutes’ whole cell lysates 15ug. Then the membrane was incubated with phosphatase. Lane 4: None, Observed MW 78 kDa, Exposure time 5 seconds

    • Primary ab dilution 1: 1000 dilution (2.355 μg /ml), Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution    1:100,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM /TBST, Lane 1: PKCd S645Pho peptides, Lane 2: PKCd unmodified peptides, Lane 3: None, Lane 4: None, Observed MW   N/A, Exposure time   3 minutes

    • Antigen PKCd S645Pho peptide, PKCd S645Pho unmodified peptide, Antigen concentration 1000ng/ml, Primary antibody concentration range 0~1000ng/ml, Secondary antibody Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), Secondary antibody concentration 1:2500 dilution

    References

    ab108972 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Permeabilization
    Yes - 0.5% Triton X100 in PBS
    Fixative
    Paraformaldehyde
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Jan 23 2014

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa Cervical Adenocarcinoma, Epithelial)
    Loading amount
    40 µg
    Specification
    HeLa Cervical Adenocarcinoma, Epithelial
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Tomar Ghansah

    Verified customer

    Submitted Feb 12 2013

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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