Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)

Overview

  • Product nameAnti-PKC alpha (phospho T497) antibody [EP2608Y]
    See all PKC alpha primary antibodies
  • Description
    Rabbit monoclonal [EP2608Y] to PKC alpha (phospho T497)
  • SpecificityDetects PKC alpha only when phosphorylated on threonine 497.
  • Tested applicationsSuitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human PKC alpha (phospho T497). The exact sequence is proprietary.
    (Peptide available as ab178734)

  • Positive control
    • WB: Jurkat cell lysate. IHC-P: Human kidney tissue.
  • General notes

    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

Applications

Our Abpromise guarantee covers the use of ab76016 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 1/50000 - 1/100000. Predicted molecular weight: 77 kDa.
IP 1/50.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notesIs unsuitable for Flow Cyt or ICC/IF.
  • 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 T497) antibody [EP2608Y] images

    • All lanes : Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016) at 1/20000 dilution

      Lane 1 : Untreated K562 (human chronic myelogenous leukemia) whole cell lysates
      Lane 2 : K562 (human chronic myelogenous leukemia) cells treated with Calyculin A whole cell lysates
      Lane 3 : K562 (human chronic myelogenous leukemia) cells were treated with Calyculin A whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size : 77 kDa
      Observed band size : 77 kDa


      Exposure time : 3 minutes

      Blocking and diluting buffer - 5% NFDM/TBST

    • ab76016, at 1/100 dilution, staining PKC alpha in human kidney by immunohistochemistry using paraffin-embedded tissue.
    • All lanes : Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016) at 1/20000 dilution

      Lane 1 : Untreated HeLa (human cervix adenocarcinoma) whole cell lysates
      Lane 2 : HeLa (human cervix adenocarcinoma) cells treated with Calyculin A whole cell lysates
      Lane 3 : HeLa (human cervix adenocarcinoma) cells were treated with Calyculin A whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size : 77 kDa
      Observed band size : 77 kDa


      Exposure time : 3 minutes

      Blocking/Diluting Buffer - 5% NFDM/TBST

    • All lanes : Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016) at 1/100000 dilution

      Lane 1 : Jurkat cell lysate - untreated
      Lane 2 : Jurkat cell lysate - treated with LP

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/1000 dilution

      Predicted band size : 77 kDa
      Observed band size : 80 kDa (why is the actual band size different from the predicted?)

      LP is Lambda Phosphatase. It is not an inducer of phosphorylation but is essential to treat the membrane (not the cells).

    • Dot blot analysis of PKC alpha (pT497) peptide (Lane 1) and PKC alpha non-phospho peptide (Lane 2) labelling PKC alpha (phospho T497) with ab76016 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    References for Anti-PKC alpha (phospho T497) antibody [EP2608Y] (ab76016)

    This product has been referenced in:
    • Heiserman JP  et al. TLR4 mutation and HSP60-induced cell death in adult mouse cardiac myocytes. Cell Stress Chaperones 20:527-35 (2015). WB ; Mouse . Read more (PubMed: 25716072) »
    • Yong SM  et al. Reduced neuronal signaling in the ageing apolipoprotein-E4 targeted replacement female mice. Sci Rep 4:6580 (2014). WB ; Mouse . Read more (PubMed: 25301084) »

    See all 5 Publications for this product

    Product Wall

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample Mouse Tissue sections (brain)
    Antigen retrieval step None
    Permeabilization No
    Specification brain
    Blocking step BSA 3% in PBS 1X + 0,5% Triton-X as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative Formaldehyde
    Username

    Dr. stefano pietropaoli

    Verified customer

    Submitted Jul 26 2016

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