Anti-PKC epsilon (phospho S729)抗体(ab63387)

概述

  • 产品名称
    Anti-PKC epsilon (phospho S729)抗体
    参阅全部 PKC epsilon 一抗
  • 描述
    兔多克隆抗体to PKC epsilon (phospho S729)
  • 特异性
    Detects endogenous levels of PKC epsilon only when phosphorylated at serine 729.
  • 经测试应用
    适用于: ICC/IF, WB, ELISA, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Guinea pig, Human
  • 免疫原

    Synthetic phosphopeptide from around the phosphorylation site of serine 729 (GFSPYF) of human PKC epsilon

  • 阳性对照
    • HeLa cells treated with PMA (125ng/ml, 30mins) Jurkat cells treated with PMA (125ng/ml, 30mins) NIH/3T3 cells treated with PMA (125ng/ml, 30mins) This antibody gave a positive result in IF in the following Formaldehyde fixed cell line: HeLa.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • 存储溶液
    Preservative: 0.02% Sodium Azide
    Constituents: 50% Glycerol, PBS (without Mg2+ and Ca2+), 150mM Sodium chloride, pH 7.4
  • Concentration information loading...
  • 纯度
    Immunogen affinity purified
  • 纯化说明
    The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
  • 克隆
    多克隆
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab63387 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
ICC/IF Use a concentration of 5 µg/ml.
WB 1/500 - 1/1000. Detects a band of approximately 84 kDa (predicted molecular weight: 84 kDa).
ELISA 1/10000.
IP Use a concentration of 5 µg/ml.

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.
  • 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 and the C2 domain is a non-calcium binding domain.
  • 翻译后修饰
    Phosphorylation on Thr-566 by PDPK1 triggers autophosphorylation on Ser-729.
  • Information by UniProt
  • 数据库链接
  • 别名
    • KPCE_HUMAN antibody
    • MGC125656 antibody
    • MGC125657 antibody
    • nPKC epsilon antibody
    • nPKC-epsilon antibody
    • PKCE antibody
    • Pkcea antibody
    • PRKCE antibody
    • Protein kinase C epsilon antibody
    • Protein kinase C epsilon type antibody
    see all

图片

  • All lanes : Anti-PKC epsilon (phospho S729) antibody (ab63387) at 1/500 dilution

    Lane 1 : HeLa cells treated with PMA (125ng/ml, 30mins) at 30 µg
    Lane 2 : Jurkat cells treated eith PMA (125ng/ml, 30mins) at 30 µg
    Lane 3 : NIH/3T3 cells treated with PMA (125ng/ml, 30mins) at 30 µg
    Lane 4 : NIH/3T3 cells treated with syntesized peptide (125ng/ml, 30mins) at 10 µg


    Predicted band size : 84 kDa
    Observed band size : 84 kDa
  • PKC epsilon was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to PKC epsilon and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab63387.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 84kDa; PKC epsilon
  • ICC/IF image of ab63387 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab63387 at 5µg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-rabbit DyLight® 488 (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

文献

This product has been referenced in:
  • Wang C  et al. Role of mitochondrial ATP-sensitive potassium channel-mediated PKC-e in delayed protection against myocardial ischemia/reperfusion injury in isolated hearts of sevoflurane-preconditioned rats. Braz J Med Biol Res 48:528-36 (2015). WB ; Rat . Read more (PubMed: 25831209) »
  • Björnström K  et al. Orexin A inhibits propofol-induced neurite retraction by a phospholipase D/protein kinase Ce-dependent mechanism in neurons. PLoS One 9:e97129 (2014). WB ; Rat . Read more (PubMed: 24828410) »

See all 2 Publications for this product

客户评价及客户问答

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing (10%)
Sample
Guinea pig Cell lysate - other (cardiac myocyte)
Specification
cardiac myocyte
Treatment
100nM drug1 for 30min, 1uM drug1 for 30min, 10uM drug1 for 30min, 100uM drug1 for 15min, 100uM drug1 for 30min, 100uM drug1 for 60min
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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提交于 Jul 07 2014

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