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)
  • Host species
    Rabbit
  • 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:
  • Temerozo JR  et al. The Neuropeptides Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide Control HIV-1 Infection in Macrophages Through Activation of Protein Kinases A and C. Front Immunol 9:1336 (2018). Read more (PubMed: 29951068) »
  • Haemmerle M  et al. Platelets reduce anoikis and promote metastasis by activating YAP1 signaling. Nat Commun 8:310 (2017). Read more (PubMed: 28827520) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Answer

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 retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins.

If there is anything else I can help you with, or if you have any issues with the new antibody, please let me know.

Read More

Answer

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 *.

If there is anything else I can help you with, or if you have any issues with the new antibody, please let me know.

Read More

Answer

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:

https://www.abcam.com/PKC-alpha-phospho-T497-antibody-EP2608Y-ab76016.html
https://www.abcam.com/PKC-alpha-phospho-T638-antibody-E195-ab32502.html



I would be more than happy to send you either of these antibodies as a replacement, just let me know which one you would like to receive. If you could also let me know either the Abcam order number or PO number that was used to purchase this antibody, then that would be very helpful to.

I look forward to your reply.

Read More

Answer

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 would like to receive and if you could provide me with the Abcam order number or the PO# that was used to purchase the antibody, that would be helpful. Or if you would prefer a refund, just let me know.

I look forward to your reply.

Read More

Question
Answer

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 are seeing with the lab testing.

I would just like to assure you that the antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples. If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

I look forward to your reply.

Read More

Answer

Codes offre de test :
ab32515 : *****
ab32096 : *****
ab7217 : *****
ab12135 : *****
ab1791 : *****

Date d’expiration : JJ MM AAAA

Je suis ravi que vous souhaitiez participer ànotre offre et tester les anticorps listés ci-dessusavec le robot Simon de Protein Simple.Ces codes de réductionsont utilisables sous 120 jours et pour l’achat d’autres anticorps primaires.

Vous devez dans un premier temps soumettre une Abreview avec vos résultats enspécifiant le code promotionnel correspondant à l'anticorps testé. Ce code sera ainsi activé et vous pourrez par la suite contacter notre service clientèle par téléphone pour passer votre prochaine commande, en vous munissant des informations suivantes : code de réduction, numéro de commande de l'anticorps testé.

Concernantles produits pour lesquels vousavez déjà obtenu des résultats, merci dem'envoyer vos données, en échange nous vous attribueronsdes"Abpoints" utilisables pour des remisessur vos commandes ouen bons d'achatAmazon.

Je reste à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview je vous souhaite unebonne continuation dans vos recherches.

Read More
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

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

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

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

Abcam user community

Verified customer

Submitted Feb 04 2011

1-10 of 11 Abreviews or Q&A

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

Sign up