Overview

  • Product name
    Anti-ROCK2 antibody [EPR7141(B)]
    See all ROCK2 primary antibodies
  • Description
    Rabbit monoclonal [EPR7141(B)] to ROCK2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ROCK2 (C terminal). The exact sequence is proprietary.

  • Positive control
    • HeLa, HEK293, L6, RAW264.7 and A673 cell lysates; Human kidney tissue, Mouse and Rat brain tissue lysate.
  • General notes

    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.

Properties

Applications

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.

Application Abreviews Notes
WB 1/10000 - 1/50000. Detects a band of approximately 161 kDa (predicted molecular weight: 161 kDa).
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Regulates the assembly of the actin cytoskeleton. Promotes formation of stress fibers and of focal adhesion complexes. Plays a role in smooth muscle contraction.
    • Sequence similarities
      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.
    • Post-translational
      modifications
      Phosphorylated upon DNA damage, probably by ATM or ATR.
    • Cellular localization
      Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • 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

    Images

    • 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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
      All lanes : 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
      All lanes : 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

    References

    This product has been referenced in:
    • Tang L  et al. RhoA/ROCK signaling regulates smooth muscle phenotypic modulation and vascular remodeling via the JNK pathway and vimentin cytoskeleton. Pharmacol Res 133:201-212 (2018). Read more (PubMed: 29791873) »
    • Vieira GM  et al. Prognostic value and functional role of ROCK2 in pediatric Ewing sarcoma. Oncol Lett 15:2296-2304 (2018). Read more (PubMed: 29434937) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    1-8 of 8 Abreviews or Q&A

    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

    Abcam user community

    Verified customer

    Submitted 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

    Abcam user community

    Verified customer

    Submitted 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

    Abcam user community

    Verified customer

    Submitted 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

    Dr. Sofia Santos

    Verified customer

    Submitted Apr 25 2013

    Answer

    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 general advise on e.g. blocking agents when we (or our customers) came along significant problems like when the antibody will work only with 3% BSA or needs 1h incubation only. Please keep in mind that every antibody needs it individual optimization, too.

    Nonetheless, for your convenience, I have attached our standard protocol to this email.



    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Answer

    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% to ROCK1. The specificity was tested performing a dotBlot.

    However, I can confirm that the immunogen sequence for ab125025 is not contained in the amino acid sequence of ROCK1 at all. This makes this product very specific for the detection of ROCK2.

    For this reason and because the ab125025 is covered by our six month guarantee for more applications (WB, IHC-P, ICC/IF) and species (Mouse, Rat, Human) I would recommend using the ab125025.

    If you want to perform IP experiments you might be eligible for the AbTrial with ab125025. You can read more about our AbTrial offer here:

    https://www.abcam.com/index.html?pageconfig=resource&rid=11998

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Answer

    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, ab58305, ab125025

    ab71598, ab66310, ab10549, ab97592, ab85612

    When selecting check the Abreview and choose the antibody which have more Abreview and publications.

    You may also interested in testing the antibody. Please click the below link for more info.

    https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    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 have 6-7 monoclonal antibodies in catalogue however we do not know whether these are being used in any functional assay. I am sorry we are unable to help with this particular ab.

    I hope this information is nevertheless helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

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

    Sign up