使用敲除细胞株进行验证RabMAb

Anti-ROCK2抗体[EPR7141(B)] (ab125025)

概述

  • 产品名称Anti-ROCK2抗体[EPR7141(B)]
    参阅全部 ROCK2 一抗
  • 描述
    兔单克隆抗体[EPR7141(B)] to ROCK2
  • 经测试应用适用于: WB, ICC/IFmore details
    不适用于: IHC-P
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human ROCK2 (C terminal).

  • 阳性对照
    • HeLa, HEK293, L6, RAW264.7 and A673 cell lysates; Human kidney tissue, Mouse and Rat brain tissue lysate.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody. 
    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.

    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.

性能

应用

Our Abpromise guarantee covers the use of ab125025 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. Detects a band of approximately 161 kDa (predicted molecular weight: 161 kDa).
ICC/IF 1/100 - 1/250.
  • 应用说明Is unsuitable for IHC-P.
  • 靶标

    • 功能Regulates the assembly of the actin cytoskeleton. Promotes formation of stress fibers and of focal adhesion complexes. Plays a role in smooth muscle contraction.
    • 序列相似性Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.
      Contains 1 AGC-kinase C-terminal domain.
      Contains 1 PH domain.
      Contains 1 phorbol-ester/DAG-type zinc finger.
      Contains 1 protein kinase domain.
      Contains 1 REM (Hr1) repeat.
    • 翻译后修饰Phosphorylated upon DNA damage, probably by ATM or ATR.
    • 细胞定位Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane.
    • Information by UniProt
    • 数据库链接
    • 别名
      • coiled-coil-containing protein kinase 2 antibody
      • KIAA0619 antibody
      • p164 ROCK 2 antibody
      • p164 ROCK-2 antibody
      • Rho associated coiled coil containing protein kinase 2 antibody
      • Rho associated protein kinase 2 antibody
      • Rho associated, coiled coil containing protein kinase II antibody
      • Rho kinase 2 antibody
      • Rho-associated antibody
      • Rho-associated protein kinase 2 antibody
      • ROCK 2 antibody
      • Rock II antibody
      • Rock2 antibody
      • ROCK2_HUMAN antibody
      • Rock2m antibody
      • ROK alpha antibody
      • ROKalpha antibody
      see all

    Anti-ROCK2 antibody [EPR7141(B)] 图像



    • Predicted band size : 161 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: ROCK2 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: A10 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab125025 observed at 165 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab125025 was shown to specifically react with ROCK2 when ROCK2 knockout samples were used. Wild-type and ROCK2 knockout samples were subjected to SDS-PAGE. ab125025 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling ROCK2 with purified ab125025 at 1/250. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291 anti-Tubulin (mouse mAb) primary and ab150120 (AlexaFluor®594 goat anti-mouse) secondary both at 1/1000 dilution. Nuclei were counterstained with DAPI (blue).

      For negative control 1, rabbit primary antibody and ab150120 (anti-mouse) secondary antibody were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by ab150077 (anti-rabbit secondary antibody).

    • All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/50000 dilution

      Lane 1 : HeLa whole cell lysate
      Lane 2 : HEK293 whole cell lysate
      Lane 3 : Mouse brain tissue lysate
      Lane 4 : Rat brain tissue lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size : 161 kDa
      Observed band size : 161 kDa
      Blocking and Diluting buffer 5% NFDM/TBST
    • All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution (unpurified)

      Lane 1 : Hela cell lysate
      Lane 2 : L6 cell lysate
      Lane 3 : A673 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size : 161 kDa
    • Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution + RAW264.7 whole cell lysate at 10 µg

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

      Predicted band size : 161 kDa
      Observed band size : 161 kDa
      Blocking and Diluting buffer 5% NFDM/TBST
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    Anti-ROCK2 antibody [EPR7141(B)] (ab125025)参考文献

    This product has been referenced in:
    • Han Y  et al. Noise-induced cochlear F-actin depolymerization is mediated via ROCK2/p-ERM signaling. J Neurochem 133:617-28 (2015). ICC/IF ; Mouse . Read more (PubMed: 25683353) »
    • Cai SD  et al. MicroRNA-144 inhibits migration and proliferation in rectal cancer by downregulating ROCK-1. Mol Med Rep 12:7396-402 (2015). Read more (PubMed: 26458302) »

    See all 2 Publications for this product

    Product Wall

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Western blot
    Sample Pig Cell lysate - whole cell (HEART)
    Gel Running Conditions Reduced Denaturing
    Loading amount 5 µg
    Specification HEART
    Blocking step Licor as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C
    Username

    Abcam user community

    Verified customer

    提交于 Nov 04 2016

    Application Western blot
    Loading amount 30 µg
    Gel Running Conditions Reduced Denaturing (12)
    Sample Human Tissue lysate - whole (CNS tissue)
    Specification CNS tissue
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Aug 14 2014

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (12)
    Sample Mouse Tissue lysate - whole (brain tissue)
    Specification brain tissue
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 May 19 2014

    Abcam has not validated the combination of species/application used in this Abreview.
    Application Immunohistochemistry (Frozen sections)
    Sample Mouse Tissue sections (Brain (hippocampus))
    Specification Brain (hippocampus)
    Fixative Paraformaldehyde
    Blocking step 10% goat serum, 2% horse serum, 1%BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
    Username

    Dr. Sofia Santos

    Verified customer

    提交于 Apr 25 2013

    Thank you for contacting us.

    As we do not beliefe in "one-fits-all" protocols, and as we do not have any advise on this datasheet, which means that most standard protocols used in our customer labs should be fine. We only give out ...

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    Thank you for contacting us and for your patience.

    I contacted the laboratory to require more information about how the specificity of ab136442 was tested. I can confirm, that the immunogen sequence of this antibody shows an identity of 75.5% ...

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    Thank you for contacting us.

    All the antibodies are well characterized, please choose based on your requirements. e.g polyclonal/ monoclonal, Rabbit/mouse/sheep etc.

    The catalogue numbers are:

    ab13660, ab45171, ab56661, a...

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    Thank you for contacting us.

    We unfortunately do not have anti ROCK antibodies available that can be used for functional assays e.g. activation or inhibition of ROCK. I would suggest searching the similar antibody clone in publications. We h...

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