Anti-PKC alpha (phospho S657 + Y658) antibody (ab23513)

Overview

  • Product nameAnti-PKC alpha (phospho S657 + Y658) antibody
    See all PKC alpha primary antibodies
  • Description
    Rabbit polyclonal to PKC alpha (phospho S657 + Y658)
  • Specificityab23513 detected PKC alpha and PKC beta I, but not other PKC isoforms in rat brain lysate.
  • Tested applicationsSuitable for: ICC/IF, WB, ELISA, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Human
  • Immunogen

    Phospho-PKC alpha (Ser-657/Tyr-658) synthetic peptide (coupled to carrier protein) corresponding to amino acid residues around serine 657 and tyrosine 658 of human PKC alpha.

  • Positive control
    • Neonatal rat brain lysate, endothelial cells, HeLa cells, rabbit spleen fibroblasts, rat pituitary cells.
  • General notesWe do not have concentration information for this antibody. It is optimized to work at the dilutions stated on the datasheet.

Properties

Applications

Our Abpromise guarantee covers the use of ab23513 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
ICC/IF Use at an assay dependent concentration. PubMed: 22532868
WB 1/1000. Detects a band of approximately 82 kDa.
ELISA 1/2000.
IP 1/100.

Use 5 - 10 µl.

IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • FunctionThis is a calcium-activated, phospholipid-dependent, serine- and threonine-specific enzyme. May play a role in cell motility by phosphorylating CSPG4.
    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.
  • Cellular localizationCytoplasm. Cell membrane. Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • AAG6 antibody
    • Aging associated gene 6 antibody
    • aPKC antibody
    • KPCA_HUMAN antibody
    • PKC alpha antibody
    • PKC-A antibody
    • PKC-alpha antibody
    • PKCA antibody
    • PRKACA antibody
    • PRKCA antibody
    • Protein Kinase C alpha antibody
    • Protein kinase C alpha type antibody
    see all

Anti-PKC alpha (phospho S657 + Y658) antibody images

  • Western blot analysis of immunoprecipitates from neonatal rat brain lysate using anti-PKCα antibody. Control and alkaline phosphatase treated precipitates were probed with anti-PKCα (Central region) or anti-phospho-PKCα (Ser-657/Tyr-658).

  • Immunocytochemical labeling of PKC phosphorylation in aldehyde-fixed and NP-40-permeabilized NGF-differentiated PC12 cells. The cells were labeled with rabbit polyclonal anti-PKCα (Ser-657/Tyr-658) (PP1091) antibody in the absence (Left) or presence (Right) of blocking peptide (PX1095). The antibody was detected using appropriate secondary antibody conjugated to DyLight® 594

  • Immunofluorescence analysis of S2-013 cells, staining PKC alpha (phospho S657 + Y658) with ab23513.

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked with blocking solution (3% BSA/PBS). Cells were incubated with primary antibody for 1 hour. An AlexaFluor®488-conjugated anti-rabbit IgG was used as the secondary antibody.
  • ab23513 staining PKC alpha (phospho S657 + Y658) in Human pancreatic ductal adenocarcinoma tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized with PBS + Triton 0.025% and blocked with 5% serum for 1 hour ta room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH 6). Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. A Biotin-conjugated Goat anti-rabbit polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

References for Anti-PKC alpha (phospho S657 + Y658) antibody (ab23513)

This product has been referenced in:
  • Wang S  et al. Regulation of endothelial cell proliferation and vascular assembly through distinct mTORC2 signaling pathways. Mol Cell Biol 35:1299-313 (2015). WB ; Mouse . Read more (PubMed: 25582201) »
  • Zhang Y  et al. Fluorosed mouse ameloblasts have increased SATB1 retention and Gaq activity. PLoS One 9:e103994 (2014). IHC-P ; Mouse . Read more (PubMed: 25090413) »

See all 5 Publications for this product

Product Wall

Thank you for your reply, and I am glad that you got the replacement antibody. We always ship our products overnight, so you can continue your research as quickly as possible.

For ab72155, we recommend starting with heat mediated antigen retri...

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Thank you for your reply.

I am sorry about the problems you have been having with ab23513 in western blot. As requested, I am sending you a vial of ab72155 as a free of charge replacement. For your records, your new order number is *.

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Thank you for your reply.

I cannot find any other antibodies that detect phosphorylated PKC on residues S657 & Y658, but we have antibodies that detected other phosphorylated residues of PKC. There are links to these below:

http:/...

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Thank you for your reply.

I am sorry that the antibody is still not working for you and so I would be happy to either replace or refund it for you.

If you would like a replacement antibody, then please let me know which antibody you w...

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Thank you for calling yesterday.

I am looking into the issues that you are having with ab23513 in western blot and I was wandering if you would be able to send an image of the results that you are getting. This will help me compare what you a...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (MDA-MB-231)
Loading amount 35 µg
Specification MDA-MB-231
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 09 2011

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 preneoplastic leasions (PanIN))
Specification mouse preneoplastic leasions (PanIN)
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate pH=6
Permeabilization Yes - PBST
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 Feb 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (mouse pancreatic cancer)
Loading amount 35 µg
Specification mouse pancreatic cancer
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 04 2011

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 (human pancreatic ductal adenocarcinoma)
Specification human pancreatic ductal adenocarcinoma
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate pH=6
Permeabilization Yes - PBS + Triton 0.025%
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 Feb 04 2011

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