使用敲除细胞株进行验证RabMAb

Anti-PKC delta (phospho S645)抗体[EP1486Y] (ab108972)

概述

  • 产品名称Anti-PKC delta (phospho S645)抗体[EP1486Y]
    参阅全部 PKC delta 一抗
  • 描述
    兔单克隆抗体[EP1486Y] to PKC delta (phospho S645)
  • 经测试应用适用于: WB, IHC-P, ICC/IFmore details
    不适用于: Flow Cyt or IP
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide corresponding to Human PKC delta aa 600 to the C-terminus (phospho S645).
    Database link: Q05655

  • 阳性对照
    • 293T cell lysate; Human breat carcinoma; HeLa cells
  • 常规说明

    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.

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

性能

应用

Our Abpromise guarantee covers the use of ab108972 in the following tested applications.

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

应用 Ab评论 说明
WB 1/10000 - 1/50000. Predicted molecular weight: 78 kDa.
IHC-P 1/50 - 1/100. Antigen retrieval is recommended.
ICC/IF 1/50 - 1/100.
  • 应用说明Is unsuitable for Flow Cyt or IP.
  • 靶标

    • 功能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.
    • 序列相似性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.
    • 结构域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.
    • 翻译后修饰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.
    • 细胞定位Cytoplasm. Membrane.
    • Information by UniProt
    • 数据库链接
    • 别名
      • 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

    Anti-PKC delta (phospho S645) antibody [EP1486Y] 图像



    • Predicted band size : 78 kDa

      Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 6 and 10: PKC delta knockout HAP1 cell lysate (20 µg)
      Lanes 3, 7 and 11: A431 cell lysate (20 µg)
      Lanes 4, 8 and 12: HeLa cell lysate (20 µg)
      Lanes 1, 2, 3 and 4: Green signal from target – ab108972 observed at 78 kDa
      Lanes 5, 6, 7 and 8: Red signal from loading control – ab8245 observed at 37 kDa
      Lanes 9, 10, 11 and 12: Merged (red and green) signal

      ab108972 was shown to specifically react with PKC delta when PKC delta knockout samples were used. Wild-type and PKC delta knockout samples were subjected to SDS-PAGE. ab108972 and ab8245 (loading control to GAPDH) were diluted 1/10000 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.



    • Predicted band size : 78 kDa

      Primary ab Dilution 1:100,000 dilution, Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution 1:20,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM/TBST, Lane 1: THP-1 whole cell lysate treated with Calyculin A with no peptides, Lane 2: THP-1 whole cell lysate treated with Calyculin A with PKCd S645Pho peptides, Lane 3: THP-1 whole cell lysate treated with Calyculin A with PKCd unmodified peptides, Lane 4: None, Observed MW   78 kDa, Exposure time   10 seconds

    • Immunofluorescent staining of PKC delta in HeLa cells, using ab108972 at a 1/50 dilution.


    • Predicted band size : 78 kDa

      Primary ab Dilution 1:100,000 dilution, Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution 1: 20,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM/TBST, Lane 1: Untreated THP-1 (Human monocytic leukemia cell line) whole cell lysates 15ug, Lane 2: THP-1 (Human monocytic leukemia cell line) treated with 50nM Calyculin A for 60 minutes whole cell lysates 15ug. Lane 3: THP-1 (Human monocytic leukemia cell line) treated with 50nM Calyculin A for 60 minutes’ whole cell lysates 15ug. Then the membrane was incubated with phosphatase. Lane 4: None, Observed MW 78 kDa, Exposure time 5 seconds

    • Immunohistochemical analysis of PKC delta in paraffin embedded Human breast carcinoma tissue, using ab108972 at a 1/50 dilution.
    • Primary ab dilution 1: 1000 dilution (2.355 μg /ml), Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution    1:100,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM /TBST, Lane 1: PKCd S645Pho peptides, Lane 2: PKCd unmodified peptides, Lane 3: None, Lane 4: None, Observed MW   N/A, Exposure time   3 minutes

    • Antigen PKCd S645Pho peptide, PKCd S645Pho unmodified peptide, Antigen concentration 1000ng/ml, Primary antibody concentration range 0~1000ng/ml, Secondary antibody Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), Secondary antibody concentration 1:2500 dilution

    Anti-PKC delta (phospho S645) antibody [EP1486Y] (ab108972)参考文献

    ab108972 has not yet been referenced specifically in any publications.

    Product Wall

    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Permeabilization Yes - 0.5% Triton X100 in PBS
    Fixative Paraformaldehyde
    Username

    Dr. Kirk Mcmanus

    Verified customer

    提交于 Jan 23 2014

    Application Western blot
    Sample Human Cell lysate - whole cell (HeLa Cervical Adenocarcinoma, Epithelial)
    Loading amount 40 µg
    Specification HeLa Cervical Adenocarcinoma, Epithelial
    Gel Running Conditions Reduced Denaturing (12%)
    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

    提交于 Feb 12 2013

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