Recombinant Anti-ROCK1 antibody [EPR638Y] (ab134181)
![Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)](https://www.abcam.com/ps/products/134/ab134181/Images/ab134181-283473-anti-rock1-antibody-epr638y-western-blot.jpg "Western blot - 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: WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-ROCK1 antibody [EPR638Y]
See all ROCK1 primary antibodies -
Description
Rabbit monoclonal [EPR638Y] to ROCK1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-Pmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
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Immunogen
Synthetic peptide within Human ROCK1 aa 1300-1400 (C terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa cell lysate, HEK293 cell lysate; IHC: Human lymphoma tissue; IP: HeLa cell lysate.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR638Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Corresponding Antibody
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Isotype control
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KO cell lysates
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Recombinant Protein
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 |
|---|---|---|
| WB | (1) |
1/500 - 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 158 kDa).
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| IP |
1/60 - 1/120.
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| IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
1/500 - 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 158 kDa). |
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IP
1/60 - 1/120. |
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IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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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
modificationsAutophosphorylated 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
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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
see all -
Alternative names
- coiled-coil-containing protein kinase 1 antibody
- coiled-coil-containing protein kinase I antibody
- MGC131603 antibody
see all
Images
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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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EPR638Y] (ab134181)](https://www.abcam.com/ps/products/134/ab134181/Images/ab134181-148500-anti-rock1-antibody-epr638y-immunohistochemistry.jpg)
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.
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![Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)](https://www.abcam.com/ps/products/134/ab134181/Images/ab134181-447961-anti-rock1-antibody-epr638y-western-blot-hela-lysates.jpg)
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.
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![Immunoprecipitation - Anti-ROCK1 antibody [EPR638Y] (ab134181)](https://www.abcam.com/ps/products/134/ab134181/Images/ab134181-6-anti-rock1-antibody-epr638y-immunoprecipitation-hela-human.jpg)
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 -
![Western blot - Anti-ROCK1 antibody [EPR638Y] (ab134181)](https://www.abcam.com/ps/products/134/ab134181/Images/ab134181-148501-anti-rock1-antibody-epr638y-western-blot.jpg)
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 -
![Anti-ROCK1 antibody [EPR638Y] (ab134181)](https://www.abcam.com/ps/products/134/ab134181/Images/ab134181-7-benefits-of-recombinant-antibodies.png)
Anti-ROCK1 antibody [EPR638Y] (ab134181)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (15)
ab134181 has been referenced in 15 publications.
- Pinto-Costa R et al. Profilin 1 delivery tunes cytoskeletal dynamics toward CNS axon regeneration. J Clin Invest 130:2024-2040 (2020). PubMed: 31945017
- 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). PubMed: 31257936
- Steurer S et al. Up regulation of Rho-associated coiled-coil containing kinase1 (ROCK1) is associated with genetic instability and poor prognosis in prostate cancer. Aging (Albany NY) 11:7859-7879 (2019). PubMed: 31557128
- Libanje F et al. ROCK2 inhibition triggers the collective invasion of colorectal adenocarcinomas. EMBO J 38:e99299 (2019). PubMed: 31304629
- 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). PubMed: 29424004