重组
RabMAb

Anti-PKC epsilon抗体[EPR1482(2)] (ab124806)

概述

  • 产品名称
    Anti-PKC epsilon抗体[EPR1482(2)]
    参阅全部 PKC epsilon 一抗
  • 描述
    兔单克隆抗体[EPR1482(2)] to PKC epsilon
  • 特异性
    85% homology with PKC alpha, beta and eta
  • 经测试应用
    适用于: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide within Human PKC epsilon aa 550-650. The exact sequence is proprietary.

  • 阳性对照
    • WB: SH-SY5Y, MCF7, HT29 and Jurkat cell lysates. Mouse and rat heart tissue lysates. IHC-P: Human kidney and endometrium tissues. ICC/IF: HeLa cells. Flow Cyt: SH-SY5Y cells.
  • 常规说明

    A trial size is available to purchase for this antibody.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

  • 形式
    Liquid
  • 存放说明
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • 解离常数(KD
    KD = 2.59 x 10 -10 M
    Learn more about KD
  • 存储溶液
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • 纯度
    Protein A purified
  • 克隆
    单克隆
  • 克隆编号
    EPR1482(2)
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab124806 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 1/1000 - 1/2000. Detects a band of approximately 82 kDa (predicted molecular weight: 84 kDa).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/30.

Flow Cyt 1/200.

For unpurified use at 1/150.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/200.

For unpurified use at 1/150.

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

  • Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1000 dilution (unpurified) + Jurkat cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PKC epsilon with purified ab124806 at 1/150. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/150) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium tissue labelling PKC epsilon with purified ab124806 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

  • Overlay histogram showing SH-SY5Y cells stained with unpurified ab124806 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab124806, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG; H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1500 dilution (purified) + Jurkat cell lysate at 10 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1000 dilution (unpurified)

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Rat heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/1500 dilution (purified)

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : Rat heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-PKC epsilon antibody [EPR1482(2)] (ab124806) at 1/10000 dilution (unpurified)

    Lane 1 : SH-SY5Y cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : HT29 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 84 kDa
    Observed band size : 82 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human endometrium tissue labelling PKC epsilon with unpurified ab124806 at 1/30. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling PKC epsilon with unpurified ab124806 at 1/100 dilution.

  • Unpurified ab124806 staining PKC epsilon in human HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol. Samples were incubated with primary antibody (1/200 in PBS) for 16 hours at 22°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counterstained with DAPI.

    See Abreview

  • Flow cytometry analysis of SH-SY5Y cells labelling PKC epsilon with purified ab124806 (red) at 1/200. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

文献

This product has been referenced in:
  • Martínez-Sánchez N  et al. Hypothalamic AMPK-ER Stress-JNK1 Axis Mediates the Central Actions of Thyroid Hormones on Energy Balance. Cell Metab 26:212-229.e12 (2017). WB ; Mouse, Rat . Read more (PubMed: 28683288) »
  • Pike T  et al. PKC? switches Aurora B specificity to exit the abscission checkpoint. Nat Commun 7:13853 (2016). ICC/IF . Read more (PubMed: 28004745) »

See all 3 Publications for this product

客户评价及客户问答

Application
Western blot
Sample
Rat Tissue lysate - other (heart)
Gel Running Conditions
Reduced Denaturing (9)
Loading amount
60 µg
Treatment
TNF-a 30min
Specification
heart
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Mr. Baptiste Jude

Verified customer

提交于 Jul 07 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
No
Fixative
Methanol
Username

Dr. Kirk Mcmanus

Verified customer

提交于 Jan 22 2014

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

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