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  1. Link

    rock1-antibody-epr638y-ab134181.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin Assembly
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ROCK1 antibody [EPR638Y] (ab134181)

  • Datasheet
  • SDS
Reviews (4)Q&A (1)References (20)

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Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)
  • Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
  • Flow Cytometry (Intracellular) - Anti-ROCK1 antibody [EPR638Y] (ab134181)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)
  • Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
  • Immunoprecipitation - Anti-ROCK1 antibody [EPR638Y] (ab134181)
  • Anti-ROCK1 antibody [EPR638Y] (ab134181)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR638Y] to ROCK1
  • Suitable for: Flow Cyt (Intra), WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

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

    Rabbit monoclonal [EPR638Y] to ROCK1
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HAP1, and HEK293 cell lysates, and Mouse brain and Rat brain tissue lysates IHC: Human testis tissue and Human breast carcinoma tissue IP: HeLa cell lysate Flow Cyt (Intra): HeLa cells
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR638Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Assembly
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Cell Biology
    • Cell Cycle
    • Cell Division
    • Centromere
    • Cancer
    • Cell cycle
    • Cell division

Associated products

  • Alternative Versions

    • Anti-ROCK1 antibody [EPR638Y] - BSA and Azide free (ab230799)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Corresponding Antibody

    • Human ROCK1 peptide (ab254529)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human ROCK1 protein (ab51415)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/150.

Not suitable for unpurified form.

WB (1)
1/500 - 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 158 kDa).
IP
1/60 - 1/120.
IHC-P
1/1200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols

For unpurified use at 1/50 - 1/100.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Notes
Flow Cyt (Intra)
1/150.

Not suitable for unpurified form.

WB
1/500 - 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 158 kDa).
IP
1/60 - 1/120.
IHC-P
1/1200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols

For unpurified use at 1/50 - 1/100.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

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.
  • Target information above from: UniProt accession Q13464 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6093 Human
    • Entrez Gene: 19877 Mouse
    • Entrez Gene: 81762 Rat
    • Omim: 601702 Human
    • SwissProt: Q13464 Human
    • SwissProt: P70335 Mouse
    • SwissProt: Q63644 Rat
    • Unigene: 306307 Human
    • Unigene: 6710 Mouse
    • Unigene: 89756 Rat
    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.

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

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling ROCK1 with Purified ab134181 at 1:1200 (1.28 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    All lanes : Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/1000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : Mouse brain
    Lane 3 : Rat brain

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 158 kDa
    Observed band size: 158 kDa

  • Flow Cytometry (Intracellular) - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    Flow Cytometry (Intracellular) - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling ROCK1 with purified ab134181 at 1/150 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150081) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (blue).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue sections labeling ROCK1 with Purified ab134181 at 1:1200 (1.28 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    All lanes : Anti-ROCK1 antibody [EPR638Y] (ab134181) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ROCK1 CRISPR/Cas9 edited HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 158 kDa
    Observed band size: 160 kDa why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab134181 observed at 160 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab134181 was shown to react with ROCK1 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264780 (CRISPR/Cas9 edited cell lysate ab257642) lane below 160kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ROCK1 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab134181 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation - Anti-ROCK1 antibody [EPR638Y] (ab134181)
    Immunoprecipitation - Anti-ROCK1 antibody [EPR638Y] (ab134181)

    Purified ab134181 at 1/120 dilution (2µg) immunoprecipitating ROCK1 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab134181 + HeLa whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134181 in HeLa whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 158 kDa

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

Protocols

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (20)

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

ab134181 has been referenced in 20 publications.

  • Zhao M  et al. MicroRNA-153 attenuates hypoxia-induced excessive proliferation and migration of pulmonary arterial smooth muscle cells by targeting ROCK1 and NFATc3. Mol Med Rep 23:N/A (2021). PubMed: 33495839
  • Morimoto A  et al. SLPI is a critical mediator that controls PTH-induced bone formation. Nat Commun 12:2136 (2021). PubMed: 33837198
  • Koizumi Y  et al. Y-27632, a ROCK inhibitor, improved laser-induced shock wave (LISW)-induced cochlear synaptopathy in mice. Mol Brain 14:105 (2021). PubMed: 34217338
  • Pinto-Costa R  et al. Profilin 1 delivery tunes cytoskeletal dynamics toward CNS axon regeneration. J Clin Invest 130:2024-2040 (2020). PubMed: 31945017
  • Koizumi Y  et al. Regenerative Effect of a ROCK Inhibitor, Y-27632, on Excitotoxic Trauma in an Organotypic Culture of the Cochlea. Front Cell Neurosci 14:572434 (2020). PubMed: 33328888
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

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

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (MDCK)
Permeabilization
Yes - 0.1% Tx-100
Specification
MDCK
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Oct 19 2020

Western blot abreview for Anti-ROCK1 antibody [EPR638Y]

Good
Abreviews
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
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

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

Good
Abreviews
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

Question

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



Thanks

Read More

Abcam community

Verified customer

Asked on Sep 27 2012

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

Abcam Scientific Support

Answered on Sep 27 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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