Overview

  • Product nameAnti-PKC antibody [MC5]
    See all PKC primary antibodies
  • Description
    Mouse monoclonal [MC5] to PKC
  • Specificitythis antibody reacts with PKC alpha, crossreactivity to PKC beta isoformes can be observed
  • Tested applicationsSuitable for: ELISA, ICC, IP, RIA, WB, Flow Cyt, IHC-FoFr, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Purified bovine brain protein kinase C

  • General notes

    Use: It should be possible to selectively block degradation of PKC without affecting the membrane associated activation. This will allow an assessment of the role of proteolysis in the activation of the protein kinase pathway.

Properties

Applications

Our Abpromise guarantee covers the use of ab31 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
ELISA Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
RIA Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.
IHC-FoFr Use at an assay dependent concentration. PubMed: 18368118
IHC-Fr 1/500.
IHC-P Use at an assay dependent concentration. PubMed: 17266784

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 antibody [MC5] images

  • Overlay histogram showing Jurkat cells stained with ab31 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab31, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse Alexa Fluor® 488 (IgG; H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HL60 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Vero cells with ab31 at a 1/50 dilution. This picture was kindly submitted as part of the review by Martha Simpson-Holley.

  • 293 T cells with ab31 at 1/50 dilution. This picture was kindly submitted as part of the review by Martha Simpson-Holley.
  • WB using ab31. Lane 1. Calpain treated PKC from Sigma . Lane 2. Homogenate from rat adipocytes. Lane 3. Homogenate from 3T3-L1 cells (mouse). Lane 4. Purified microsomes from 3T3-L1 cells. Review by Mikael Rutberg submitted 2 September 2004.

    WB using ab31. Lane 1. Calpain treated PKC from Sigma . Lane 2. Homogenate from rat adipocytes. Lane 3. Homogenate from 3T3-L1 cells (mouse). Lane 4. Purified microsomes from 3T3-L1 cells. Review by Mikael Rutberg submitted 2 September 2004.
  • ab31 staining PKC in Mouse spleen and duodenum tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/500 in PBS) for 8 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody. Nuclei were stained by DAPI.

    See Abreview

  • ab31 staining PKC in Human skin papilloma tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

References for Anti-PKC antibody [MC5] (ab31)

This product has been referenced in:
  • Samuel MA  et al. LKB1 and AMPK regulate synaptic remodeling in old age. Nat Neurosci 17:1190-7 (2014). IHC-FoFr, IHC - Wholemount ; Mouse . Read more (PubMed: 25086610) »
  • De la Huerta I  et al. Direction-selective retinal ganglion cells arise from molecularly specified multipotential progenitors. Proc Natl Acad Sci U S A 109:17663-8 (2012). IHC-FoFr ; Mouse . Read more (PubMed: 23045641) »

See all 12 Publications for this product

Product Wall

Indeed, this antibody is a clone and is used by many researchers. However I could not conclude on its specificity. Indeed, some sources quote that this antibody is specific for PKC alpha, whereas other state that there might be crossreactivity with PKC...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (human pancreatic cancer cell lines)
Loading amount 30 µg
Specification human pancreatic cancer cell lines
Gel Running Conditions Non-reduced Denaturing (7.5%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Dr. Pawel Mazur

Verified customer

Submitted Jul 31 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Skin papiloma)
Specification Skin papiloma
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 15 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (mouse pancreatic neoplasia)
Specification mouse pancreatic neoplasia
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization No
Blocking step MOM kit blocking agaent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 14 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (pancreatic tisse and tumor)
Specification pancreatic tisse and tumor
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Mouse spleen and duodenum)
Specification Mouse spleen and duodenum
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Pancreatic cancer)
Loading amount 35 µg
Specification Pancreatic cancer
Gel Running Conditions Non-reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Dr. Pawel Mazur

Verified customer

Submitted Jun 04 2010

Thank you for your enquiry. For ab4127, this antibody was validated on tPA induced myeloid cells for IHC. Also, melanocytes have been known to express PKC alpha and alternatively brain tissue is typically inherently expressive. For ab31, I regret t...

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I apologize for the delay in responding to your enquiry. The originator of this antibody attests that this antibody has been found to recognize Protein kinase Ca. Epitope: within the variable region V3 PKCa (the site of limited proteolysis on the nativ...

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We have no information about the dilutions originally used with this antibody. I would suggest trying the antibody at 1:1000, 1:250 and 1:2000 initially, the ideal dilution will depend on the details of your experiments.

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