Recombinant
RabMAb

Anti-PKC antibody [EPR16794] (ab179521)

Overview

  • Product name
    Anti-PKC antibody [EPR16794]
    See all PKC primary antibodies
  • Description
    Rabbit monoclonal [EPR16794] to PKC
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human PKC alpha+beta1+beta2 aa 450 to the C-terminus. The exact sequence is proprietary.
    Database link: P05771

  • Positive control
    • WB: Active human PKC alpha full length protein; Active human PKC beta 1 full length protein; Active human PKC beta 2 full length protein; Active human PKC gamma full length protein; Human fetal brain and fetal kidney lysates; Jurkat, SH-SY5Y and L-929 whole cell lysates; Rat and mouse thymus lysates. IHC-P: Human breast cancer, Mouse liver and Rat testis tissues. ICC/IF: HeLa cells. IP: Mouse brain whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

    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

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR16794
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab179521 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 1/2000. Detects a band of approximately 78 kDa (predicted molecular weight: 77, 78 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/80.

Target

  • Function
    Calcium-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 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. 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

Images

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

    Predicted band size : 77, 78 kDa


    Exposure time : 5 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

    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 gamma full length protein (Catalog#ab60842) contains aa1-677 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.

  • 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

    Blocking/Dilution buffer: 5% NFDM/TBST.

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

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

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

  • 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
    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?)

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-PKC antibody [EPR16794] (ab179521) at 1/5000 dilution

    Lane 1 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
    Lane 2 : Rat thymus lysate
    Lane 3 : Mouse thymus lysate

    Lysates/proteins at 10 µg per lane.

    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?)

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

References

ab179521 has not yet been referenced specifically in any publications.

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