Overview

  • Product nameAnti-PKC (phospho T514) antibody [EP2730Y]
    See all PKC primary antibodies
  • Description
    Rabbit monoclonal [EP2730Y] to PKC (phospho T514)
  • Tested applicationsSuitable for: Dot blot, WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    corresponding to Human PKC (phospho T514).

  • Positive control
    • HeLa cell lysate; Human brain tissue
  • General notes

    A trial size is available to purchase for this antibody.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109539 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
Dot blot 1/1000.
WB 1/1000 - 1/10000. Predicted molecular weight: 78 kDa.
IP 1/50.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notesIs unsuitable for Flow Cyt or ICC/IF.
  • Target

    • FunctionCalcium-activated, phospholipid- and diacylglycerol (DAG)-dependent serine/threonine-protein kinase that is involved in positive and negative regulation of cell proliferation, apoptosis, differentiation, migration and adhesion, tumorigenesis, cardiac hypertrophy, angiogenesis, platelet function and inflammation, by directly phosphorylating targets such as RAF1, BCL2, CSPG4, TNNT2/CTNT, or activating signaling cascade involving MAPK1/3 (ERK1/2) and RAP1GAP. Involved in cell proliferation and cell growth arrest by positive and negative regulation of the cell cycle. Can promote cell growth by phosphorylating and activating RAF1, which mediates the activation of the MAPK/ERK signaling cascade, and/or by up-regulating CDKN1A, which facilitates active cyclin-dependent kinase (CDK) complex formation in glioma cells. In intestinal cells stimulated by the phorbol ester PMA, can trigger a cell cycle arrest program which is associated with the accumulation of the hyper-phosphorylated growth-suppressive form of RB1 and induction of the CDK inhibitors CDKN1A and CDKN1B. Exhibits anti-apoptotic function in glioma cells and protects them from apoptosis by suppressing the p53/TP53-mediated activation of IGFBP3, and in leukemia cells mediates anti-apoptotic action by phosphorylating BCL2. During macrophage differentiation induced by macrophage colony-stimulating factor (CSF1), is translocated to the nucleus and is associated with macrophage development. After wounding, translocates from focal contacts to lamellipodia and participates in the modulation of desmosomal adhesion. Plays a role in cell motility by phosphorylating CSPG4, which induces association of CSPG4 with extensive lamellipodia at the cell periphery and polarization of the cell accompanied by increases in cell motility. Is highly expressed in a number of cancer cells where it can act as a tumor promoter and is implicated in malignant phenotypes of several tumors such as gliomas and breast cancers. Negatively regulates myocardial contractility and positively regulates angiogenesis, platelet aggregation and thrombus formation in arteries. Mediates hypertrophic growth of neonatal cardiomyocytes, in part through a MAPK1/3 (ERK1/2)-dependent signaling pathway, and upon PMA treatment, is required to induce cardiomyocyte hypertrophy up to heart failure and death, by increasing protein synthesis, protein-DNA ratio and cell surface area. Regulates cardiomyocyte function by phosphorylating cardiac troponin T (TNNT2/CTNT), which induces significant reduction in actomyosin ATPase activity, myofilament calcium sensitivity and myocardial contractility. In angiogenesis, is required for full endothelial cell migration, adhesion to vitronectin (VTN), and vascular endothelial growth factor A (VEGFA)-dependent regulation of kinase activation and vascular tube formation. Involved in the stabilization of VEGFA mRNA at post-transcriptional level and mediates VEGFA-induced cell proliferation. In the regulation of calcium-induced platelet aggregation, mediates signals from the CD36/GP4 receptor for granule release, and activates the integrin heterodimer ITGA2B-ITGB3 through the RAP1GAP pathway for adhesion. During response to lipopolysaccharides (LPS), may regulate selective LPS-induced macrophage functions involved in host defense and inflammation. But in some inflammatory responses, may negatively regulate NF-kappa-B-induced genes, through IL1A-dependent induction of NF-kappa-B inhibitor alpha (NFKBIA/IKBA). Upon stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA), phosphorylates EIF4G1, which modulates EIF4G1 binding to MKNK1 and may be involved in the regulation of EIF4E phosphorylation. Phosphorylates KIT, leading to inhibition of KIT activity. Phosphorylates ATF2 which promotes cooperation between ATF2 and JUN, activating transcription.
    • Sequence similaritiesBelongs 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 localizationCytoplasm. Cell membrane. Mitochondrion membrane. Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • KPCA_HUMAN antibody
      • PKC alpha antibody
      • PKC beta antibody
      • PKC delta antibody
      • PKC epsilon antibody
      • PKC gamma antibody
      • PKC zeta antibody
      • PKC-A antibody
      • PKC-alpha antibody
      • PKC2 antibody
      • PKCA antibody
      • PKCB antibody
      • PKCD antibody
      • PKCE antibody
      • PKCG antibody
      • PRKCA antibody
      • PRKCB antibody
      • PRKCD antibody
      • PRKCE antibody
      • PRKCG antibody
      • PRKCZ antibody
      • Protein kinase C alpha antibody
      • Protein kinase C alpha type antibody
      • Protein kinase C beta antibody
      • Protein kinase C delta antibody
      • Protein kinase C epsilon antibody
      • Protein kinase C gamma antibody
      • Protein kinase C zeta antibody
      see all

    Anti-PKC (phospho T514) antibody [EP2730Y] images

    • All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) 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 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
      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

      Blocking and Diluting buffer and concentration

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

    • All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) at 1/5000 dilution

      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
      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

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Dot blot analysis of PKC (pT514) phospho peptide (Lane 1) and PKC non-phospho peptide (Lane 2) using ab109539 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

      Blocking and Diluting buffer and concentration: 5% NFDM /TBST.

      Exposure time: 3 minutes.

    • All lanes : Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539) at 0.02 µg/ml

      Lane 1 : HeLa cell lysate with PKC gamma (pT514) peptide
      Lane 2 : HeLa cell lysate with PKC gamma non-phospho peptide
      Lane 3 : 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
    • ab109539 at 1/100 dilution staining PKC (phospho T514) in paraffin-embedded Human brain tissue by Immunohistochemistry.

    References for Anti-PKC (phospho T514) antibody [EP2730Y] (ab109539)

    This product has been referenced in:
    • Huang L  et al. Ketamine potentiates hippocampal neurodegeneration and persistent learning and memory impairment through the PKC?-ERK signaling pathway in the developing brain. Brain Res 1476:164-71 (2012). WB . Read more (PubMed: 22985497) »
    • Dunn EF & Connor JH Dominant inhibition of Akt/protein kinase B signaling by the matrix protein of a negative-strand RNA virus. J Virol 85:422-31 (2011). Read more (PubMed: 20980511) »

    See all 2 Publications for this product

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