Overview

  • Product nameAnti-PKC antibody [MC5]
    See all PKC primary antibodies
  • Description
    Mouse monoclonal [MC5] to PKC
  • Conjugation notes-
  • Specificitythis antibody reacts with PKC alpha, crossreactivity to PKC beta isoformes can be observed
  • Tested applicationsELISA, 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-epitope mapping within the hinge region of PKC (residues 292-317).-


    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

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 Alexa Fluor® 488 goat anti-mouse 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:
  • 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) »
  • McMains E  et al. Expression and Localization of CLC Chloride Transport Proteins in the Avian Retina. PLoS One 6:e17647 (2011). ICC/IF ; Chicken . Read more (PubMed: 21408174) »

See all 11 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"