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

Overview

  • Product name
    Anti-PKC alpha (phospho S657 + Y658) antibody
    See all PKC alpha primary antibodies
  • Description
    Rabbit polyclonal to PKC alpha (phospho S657 + Y658)
  • Specificity
    ab23513 detected PKC alpha and PKC beta I, but not other PKC isoforms in rat brain lysate.
  • Tested applications
    Suitable 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 notes
    We 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

  • Function
    This 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 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. 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

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).

  • 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 at 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

  • 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.

References

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) »
  • Li XC  et al. Role of the Na+/H+ exchanger 3 in angiotensin II-induced hypertension in NHE3-deficient mice with transgenic rescue of NHE3 in small intestines. Physiol Rep 3:N/A (2015). WB . Read more (PubMed: 26564064) »

See all 7 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

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

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

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