Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-ROCK1 antibody [EPR638Y] (ab134181)

Reviews (3)Q&A (1)References (12)

Overview

  • Product name

    Anti-ROCK1 antibody [EPR638Y]
    See all ROCK1 primary antibodies

  • Description

    Rabbit monoclonal [EPR638Y] to ROCK1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, IHC-Fr, WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human ROCK1 aa 1300-1400 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HeLa cell lysate, HEK293 cell lysate IHC: Human lymphoma tissue.

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab134181 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
ICC/IF 1/50 - 1/100.
IHC-Fr Use at an assay dependent concentration. PubMed: 24465504
WB 1/500 - 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 158 kDa).
IP 1/60.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt.

    • Target

    • Function

      Protein kinase which is a key regulator of actin cytoskeleton and cell polarity. Involved in regulation of smooth muscle contraction, actin cytoskeleton organization, stress fiber and focal adhesion formation, neurite retraction, cell adhesion and motility via phosphorylation of DAPK3, GFAP, LIMK1, LIMK2, MYL9/MLC2, PFN1 and PPP1R12A. Phosphorylates FHOD1 and acts synergistically with it to promote SRC-dependent non-apoptotic plasma membrane blebbing. Phosphorylates JIP3 and regulates the recruitment of JNK to JIP3 upon UVB-induced stress. Acts as a suppressor of inflammatory cell migration by regulating PTEN phosphorylation and stability. Acts as a negative regulator of VEGF-induced angiogenic endothelial cell activation. Required for centrosome positioning and centrosome-dependent exit from mitosis. Plays a role in terminal erythroid differentiation. May regulate closure of the eyelids and ventral body wall by inducing the assembly of actomyosin bundles. Promotes keratinocyte terminal differentiation. Involved in osteoblast compaction through the fibronectin fibrillogenesis cell-mediated matrix assembly process, essential for osteoblast mineralization.

    • Tissue specificity

      Detected in blood platelets.

    • 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.

    • Domain

      The C-terminal auto-inhibitory domain interferes with kinase activity. RHOA binding leads to a conformation change and activation of the kinase. Truncated ROCK1 is constitutively activated.

    • Post-translational
      modifications

      Autophosphorylated on serine and threonine residues.
      Cleaved by caspase-3 during apoptosis. This leads to constitutive activation of the kinase and membrane blebbing.

    • Cellular localization

      Cytoplasm. Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, centriole. Golgi apparatus membrane. Cell projection, bleb. Cytoplasm, cytoskeleton. Cell membrane. Cell projection, lamellipodium. Cell projection, ruffle. Associated with the mother centriole and an intercentriolar linker. Colocalizes with ITGB1BP1 and ITGB1 at the cell membrane predominantly in lamellipodia and membrane ruffles, but also in retraction fibers. Localizes at the cell membrane in an ITGB1BP1-dependent manner (By similarity). A small proportion is associated with Golgi membranes.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • coiled-coil-containing protein kinase 1 antibody
      • coiled-coil-containing protein kinase I antibody
      • MGC131603 antibody
      • MGC43611 antibody
      • p160 Rhoassociated coiled coil-forming protein kinase antibody
      • p160 ROCK-1 antibody
      • p160 ROCK1 antibody
      • p160ROCK antibody
      • PRO0435 antibody
      • Renal carcinoma antigen NY REN 35 antibody
      • Renal carcinoma antigen NY-REN-35 antibody
      • Rho associated coiled coil containing protein kinase 1 antibody
      • Rho associated protein kinase 1 antibody
      • Rho kinase antibody
      • Rho-alpha kinase antibody
      • Rho-associated antibody
      • Rho-associated protein kinase 1 antibody
      • ROCK I antibody
      • ROCK-I antibody
      • ROCK1 antibody
      • ROCK1_HUMAN antibody
      • Rok antibody
      • rokalpha antibody
      see all

    Images

    • Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
      Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)

      Lane 1: Wild type HAP1 whole cell lysate (40 µg)
      Lane 2: ROCK1 knockout HAP1 whole cell lysate (40 µg)
      Lane 3: HEK293 whole cell lysate (20 µg)
      Lane 4: HeLa whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab134181 observed at 165 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab134181 was shown to specifically react with ROCK1 when ROCK1 knockout samples were used. Wild-type and ROCK1 knockout samples were subjected to SDS-PAGE. Ab134181 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-ROCK1 antibody [EPR638Y] (ab134181)
      Immunocytochemistry/ Immunofluorescence - Anti-ROCK1 antibody [EPR638Y] (ab134181)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
      ab134181 (1/200) staining ROCK1 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

      See Abreview

    • Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
      Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
      Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/500 dilution + HeLa cell lysate at 10 µg

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

      Predicted band size: 158 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)

      Immunohistochemical analysis of paraffin embedded Human lymphoma tissue labelling ROCK1 with ab134181 at a dilution of 1/50.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Jia L  et al. Shikonin inhibits proliferation, migration, invasion and promotes apoptosis in NCI-N87 cells via inhibition of PI3K/AKT signal pathway. Artif Cells Nanomed Biotechnol 47:2662-2669 (2019). Read more (PubMed: 31257936) »
    • Zhao D  et al. Substrate stiffness regulated migration and invasion ability of adenoid cystic carcinoma cells via RhoA/ROCK pathway. Cell Prolif 51:e12442 (2018). Read more (PubMed: 29424004) »
    See all 12 Publications for this product

    Publishing research using ab134181? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-ROCK1 antibody [EPR638Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (12%)
    Sample
    Rat Tissue lysate - whole (Brain)
    Specification
    Brain
    Treatment
    brain trauma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Dr. Da Liu

    Verified customer

    Submitted Mar 13 2014

    Immunohistochemistry (Frozen sections) abreview for Anti-ROCK1 antibody [EPR638Y]

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 21°C
    Sample
    Mouse Tissue sections (Choroid plexus)
    Specification
    Choroid plexus
    Permeabilization
    Yes - 0.25% Triton
    Fixative
    Paraformaldehyde
    Read More

    Dr. Sofia Santos

    Verified customer

    Submitted Oct 08 2013

    Question

    Hi I understand, so can we try the different antibody ab143181 please.



    Thanks

    Read More
    Answer

    Thank you for confirming these details and for your cooperation.

    As requested, I have issued a free of charge replacement of alternative ab134181 Anti-ROCK1 antibody [EPR638Y]with the order number #####.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to letme know. I sincerely hope this vial will work in your experiments. If not, please do not hesitate to contact me and I suggest in this case the best way to proceed would be to provide a refund or credit note for the original antibody.

    I wish you the best of luck and look forward to hearing from you with the results of the experiments with the new antibody.

    Read More

    Immunocytochemistry/ Immunofluorescence abreview for Anti-ROCK1 antibody [EPR638Y]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% Triton-X100 in PBS
    Read More

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Sep 05 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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