RecombinantRabMAb

Recombinant Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
  • Western blot - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
  • Immunoprecipitation - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
  • Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22277] to PKC theta/PRKCQ
  • Suitable for: WB, IHC-P, IP
  • Reacts with: Human

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

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)

        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).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)

        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).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)

        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).

      • Western blot - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
        Western blot - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
        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.

      • Immunoprecipitation - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
        Immunoprecipitation - Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)

        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.

      • Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)
        Anti-PKC theta/PRKCQ antibody [EPR22277] (ab230971)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab230971? Please let us know so that we can cite the reference in this datasheet.

      ab230971 has not yet been referenced specifically in any publications.

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