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RabMAb

Anti-PKC delta (phospho Y311) 抗体 [EPR2609Y] (ab76181)

製品の概要

  • 製品名
    Anti-PKC delta (phospho Y311) antibody [EPR2609Y]
    PKC delta 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR2609Y] to PKC delta (phospho Y311)
  • 特異性
    ab76181 only detects PKC delta phosphorylated on Tyrosine 311.
  • アプリケーション
    適用あり: Dot blot, WB, IP, IHC-Pmore details
    適用なし: Flow Cyt or ICC/IF
  • 種交差性
    交差種: Mouse, Human
  • 免疫原

    A synthetic phospho-peptide from residues surrounding Tyrosine 311 of human PKC delta.

  • ポジティブ・コントロール
    • WB: HeLa cell lysate treated with Phorbol-12-myristate-13-acetate and U937 cell lysate treated with TPA. IHC-P: Human urinary bladder carcinoma tissue. IP: HeLa treated with 10mM H2O2 for 1 hour whole cell lysate.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

製品の特性

関連製品

アプリケーション

Our Abpromise guarantee covers the use of ab76181 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Dot blot 1/1000.
WB 1/2500 - 1/5000. Predicted molecular weight: 77 kDa.
IP 1/40.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • 追加情報
    Is unsuitable for Flow Cyt or ICC/IF.
  • ターゲット情報

    • 機能
      This is calcium-independent, phospholipid-dependent, serine- and threonine-specific enzyme. 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. May play a role in antigen-dependent control of B-cell function. Phosphorylates MUC1 in the C-terminal and regulates the interaction between MUC1 and beta-catenin.
    • 配列類似性
      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.
    • ドメイン
      The C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor.
      The C2 domain is a non-calcium binding domain. It binds proteins containing phosphotyrosine in a sequence-specific manner.
    • 翻訳後修飾
      Phosphorylated on Thr-507, within the activation loop. Autophosphorylated and/or phosphorylated. Although the Thr-507 phosphorylation occurs it is not a prerequisite for enzymatic activity.
    • 細胞内局在
      Cytoplasm. Membrane.
    • Information by UniProt
    • 参照データベース
    • 別名
      • CVID9 antibody
      • D14Ertd420e antibody
      • Kinase PKC delta antibody
      • KPCD antibody
      • KPCD_HUMAN antibody
      • MAY 1 antibody
      • MAY1 antibody
      • MGC49908 antibody
      • nPKC delta antibody
      • nPKC-delta antibody
      • PCKd antibody
      • PKC d antibody
      • PKC delta antibody
      • PKCD antibody
      • PKCdelta antibody
      • PRKC D antibody
      • PRKC delta antibody
      • Prkcd antibody
      • Protein Kinase C delta antibody
      • Protein kinase C delta type antibody
      • Protein kinase C delta VIII antibody
      • Protein Kinase Cdelta antibody
      • Tyrosine protein kinase PRKCD antibody
      see all

    画像

    • All lanes : Anti-PKC delta (phospho Y311) antibody [EPR2609Y] (ab76181) at 1/500 dilution

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Phorbol-12-myristate-13-acetate at 200nM for 20 minutes. Whole cell lysates
      Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with Phorbol-12-myristate-13-acetate at 200nM for 20 minutes. Whole cell lysates. Then the membrane was incubated with phosphatase.

      Lysates/proteins at 15 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 77 kDa
      Observed band size : 78 kDa (why is the actual band size different from the predicted?)


      Exposure time : 3 minutes

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    • Dot blot analysis of PKC delta (pY311) phospho peptide (Lane 1) and PKC delta non-phospho peptide (Lane 2) using ab76181 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution.

      Blocking and Diluting buffer and concentration: 5% NFDM /TBST.

      Exposure time: 3 minutes.

    • Immunohistochemical analysis of paraffin-embedded human urinary bladder carcinoma with ab76181 at 1/100 dilution.
    • ab76181 at 1/20 immunoprecipitating PKC delta (phospho Y311) in HeLa treated with 10mM H2O2 for 1 hour whole cell lysate.

      Lane 1 (input): HeLa treated with 10mM H2O2 for 1 hour whole cell lysate (10µg)
      Lane 2 (+): ab76181 + HeLa treated with 10mM H2O2 for 1 hour whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76181 in HeLa treated with 10mM H2O2 for 1 hour whole cell lysate.

      For western blotting, ab76181 was used as a dilution of 1/200 followed by ab131366 VeriBlot for IP (HRP)  at a dilution of 1/1000.

      Blocking buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-PKC delta (phospho Y311) antibody [EPR2609Y] (ab76181) at 1/5000 dilution

      Lane 1 : U937 cell lysate
      Lane 2 : U937 cell lysate treated with TPA

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat anti-rabbit-HRP at 1/2000 dilution

      Predicted band size : 77 kDa
      Observed band size : 78 kDa (why is the actual band size different from the predicted?)
    • Anti-PKC delta (phospho Y311) antibody [EPR2609Y] (ab76181) at 1/2500 dilution + C57BL/6 Mouse splenocytes whole cell lysate at 40 µg

      Secondary
      Goat Anti-rabbit IgG (H&L) HRP conjugated polyclonal antibody at 1/10000 dilution

      Performed under reducing conditions.

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


      Exposure time : 10 seconds

      This image is courtesy of an abreview submitted by Bryan Niedenberger Tomar Ghansah

      Blocking- 5% Milk for 1 hour at 25°C.

      A non-specific band was observed at 80 kDa.  

      See Abreview

    参考文献

    This product has been referenced in:
    • Gruosso T  et al. Chronic oxidative stress promotes H2AX protein degradation and enhances chemosensitivity in breast cancer patients. EMBO Mol Med 8:527-49 (2016). WB . Read more (PubMed: 27006338) »
    • Ficarro SB  et al. Online nanoflow multidimensional fractionation for high efficiency phosphopeptide analysis. Mol Cell Proteomics 10:O111.011064 (2011). WB ; Human . Read more (PubMed: 21788404) »

    See all 2 Publications for this product

    レビューと Q&A

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa Cervical Adenocarcinoma, Epithelial)
    Loading amount
    40 µg
    Specification
    HeLa Cervical Adenocarcinoma, Epithelial
    Treatment
    100nM Insulin
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Tomar Ghansah

    Verified customer

    投稿 Feb 20 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (C57BL/6 Mouse splenocytes)
    Loading amount
    40 µg
    Specification
    C57BL/6 Mouse splenocytes
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Tomar Ghansah

    Verified customer

    投稿 Oct 09 2012

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

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