Overview

  • Product nameAnti-PKC epsilon antibody [EPR1482(2)]
    See all PKC epsilon primary antibodies
  • Description
    Rabbit monoclonal [EPR1482(2)] to PKC epsilon
  • Specificity85% homology with PKC alpha, beta and eta
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PKC epsilon.

  • Positive control
    • WB: SH-SY5Y, MCF7, HT29 and Jurkat cell lysates. Mouse and rat heart tissue lysates. IHC-P: Human kidney and endometrium tissues. ICC/IF: HeLa cells. Flow Cyt: SH-SY5Y cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    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.

    A trial size is available to purchase for this antibody.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD) KD = 2.59 x 10 -10 M Learn more about KD
  • Storage bufferpH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • PurityProtein A purified
  • ClonalityMonoclonal
  • Clone numberEPR1482(2)
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab124806 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 1/1000 - 1/2000. Detects a band of approximately 82 kDa (predicted molecular weight: 84 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/30.

Flow Cyt 1/200.

For unpurified use at 1/150.





ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/200.

For unpurified use at 1/150.

Target

  • FunctionThis 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.
  • 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.
  • DomainThe C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor and the C2 domain is a non-calcium binding domain.
  • Post-translational
    modifications
    Phosphorylation on Thr-566 by PDPK1 triggers autophosphorylation on Ser-729.
  • Information by UniProt
  • Database links
  • Alternative names
    • KPCE_HUMAN antibody
    • MGC125656 antibody
    • MGC125657 antibody
    • nPKC epsilon antibody
    • nPKC-epsilon antibody
    • PKCE antibody
    • Pkcea antibody
    • PRKCE antibody
    • Protein kinase C epsilon antibody
    • Protein kinase C epsilon type antibody
    see all

Anti-PKC epsilon antibody [EPR1482(2)] images

  • Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1000 dilution (unpurified) + Jurkat cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1500 dilution (purified) + Jurkat cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1000 dilution (unpurified)

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Rat heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1500 dilution (purified)

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Rat heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/10000 dilution (unpurified)

    Lane 1 : SH-SY5Y cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : HT29 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium tissue labelling PKC epsilon with unpurified ab124806 at 1/30. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium tissue labelling PKC epsilon with purified ab124806 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling PKC epsilon with unpurified ab124806 at 1/100 dilution.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PKC epsilon with purified ab124806 at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/150) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

  • Unpurified ab124806 staining PKC epsilon in human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol. Samples were incubated with primary antibody (1/200 in PBS) for 16 hours at 22°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.

    See Abreview

  • Overlay histogram showing SH-SY5Y cells stained with unpurified ab124806 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab124806, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG; H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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 SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Flow cytometry analysis of SH-SY5Y cells labelling PKC epsilon with purified ab124806 (red) at 1/200. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-PKC epsilon antibody [EPR1482(2)] (ab124806)

This product has been referenced in:

See 1 Publication for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - other (heart)
Gel Running Conditions Reduced Denaturing (9)
Loading amount 60 µg
Treatment TNF-a 30min
Specification heart
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Mr. Baptiste Jude

Verified customer

Submitted Jul 07 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization No
Fixative Methanol
Username

Dr. Kirk McManus

Verified customer

Submitted Jan 22 2014

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