Overview

  • Product name

    Anti-PKC theta/PRKCQ antibody [EPR22277]
    See all PKC theta/PRKCQ primary antibodies
  • Description

    Rabbit monoclonal [EPR22277] to PKC theta/PRKCQ
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IPmore details
    Unsuitable for: Flow Cyt or ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human PKC theta/PRKCQ aa 650 to the C-terminus. The exact sequence is proprietary.
    Database link: Q04759

  • Positive control

    • WB: Jurkat, MOLT-4 and K562 whole cell lysate. IHC-P: Human tonsil tissue. Human gastrointestinal stromal tumor (GIST). IP: PKC theta/PRKCQ IP in K562 whole cell lysate.
  • General notes

     This product was previously labelled as PKC theta

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab230971 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 1/1000. Predicted molecular weight: 82 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/30.
  • Application notes
    Is unsuitable for Flow Cyt or ICC/IF.
  • Target

    • Function

      This is a calcium-independent, phospholipid-dependent, serine- and threonine-specific enzyme. Essential for T-cell receptor (TCR)-mediated T-cell activation, but is dispensable during TCR-dependent thymocyte development. Links the TCR signaling complex to the activation of NF-kappa-B in mature T lymphocytes. Required for interleukin-2 (IL2) production.
      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.
    • Tissue specificity

      Skeletal muscle, megakaryoblastic cells and platelets.
    • 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.
    • Domain

      The C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor and the C2 domain is a non-calcium binding domain.
    • Post-translational
      modifications

      Autophosphorylation at Thr-219 is required for targeting to the TCR and cellular function of PKC upon antigen receptor ligation.
    • Information by UniProt
    • Database links

    • Alternative names

      • KPCT_HUMAN antibody
      • MGC126514 antibody
      • MGC141919 antibody
      • nPKC theta antibody
      • nPKC-theta antibody
      • nPKCtheta antibody
      • OTTHUMP00000043364 antibody
      • OTTHUMP00000043365 antibody
      • PKC 0 antibody
      • PKC0 antibody
      • Prkcq antibody
      • PRKCT antibody
      • Protein kinase C theta antibody
      • Protein kinase C theta type antibody
      • Protein Kinase Ctheta antibody
      see all

    Images

    • Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor (GIST) tissue labeling PKC theta/PRKCQ with ab230971 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining in human gastrointestinal stromal tumor (PMID: 15289315) is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PKC theta/PRKCQ with ab230971 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic and nuclear staining in T lymphocytes in tonsil (PMID: 27149922) is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemical analysis of paraffin-embedded human leiomyosarcoma tissue labeling PKC theta/PRKCQ with ab230971 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Negative control: No staining in human leiomyosarcoma (PMID: 15289315).

      Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • All lanes : Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971) at 1/1000 dilution

      Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
      Lane 2 : MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate
      Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 82 kDa
      Observed band size: 75, 40 kDa
      why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in the literature. The 40kDa band is a cleaved form (PMID: 9252332, PMID: 24924627).

      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lane 1: 15 seconds; Lanes 2-3: 3 minutes.

    • PKC theta/PRKCQ was immunoprecipitated from 0.35 mg K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab230971 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230971 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: K562 whole cell lysate 10 µg (Input).
      Lane 2: ab230971 IP in K562 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230971 in K562 whole cell lysate.

      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 3 seconds.

    References

    ab230971 has not yet been referenced specifically in any publications.

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