Anti-ROCK1 antibody [EP786Y] (ab45171)
- Datasheet
- Specific References (32)
- Protocols
Overview
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Product nameAnti-ROCK1 antibody [EP786Y]
See all ROCK1 primary antibodies -
DescriptionRabbit monoclonal [EP786Y] to ROCK1
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Host speciesRabbit
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SpecificityThis antibody recognizes both the cleaved C-terminus of ROCK 1 (30 kDa) and full length protein (158 kDa). The immunogen used for this product shares 83% homology with ROCK2. Cross-reactivity with this protein has not been confirmed experimentally.
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Tested applicationsSuitable for: IHC-Fr, WB, IHC-P, ICC/IF, IP, Flow Cytmore details
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Species reactivityReacts with: Mouse, Rat, Cow, Human
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Immunogen
Synthetic peptide within Human ROCK1 aa 1100-1200 (C terminal). The exact sequence is proprietary.
Database link: Q13464 -
Positive control
- WB: Untreated, Calyculin A treated and Camptothecin treated HeLa cell lysates. Jurkat, Ramos, PC-12 and RAW264.7 cell lysates. IHC-P: Human adenocarcinoma of the colon and thyroid gland carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa cell lysate.
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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
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEP786Y
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Positive Controls
Applications
Our Abpromise guarantee covers the use of ab45171 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 |
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| IHC-Fr | 1/100. PubMed: 19295659 | |
| WB | 1/2000 - 1/10000. Predicted molecular weight: 158 kDa. For unpurified use at 1/500. |
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| IHC-P | 1/50 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| ICC/IF | 1/50 - 1/500.
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| IP | 1/40 - 1/50.
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| Flow Cyt | 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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FunctionProtein 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.
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Tissue specificityDetected in blood platelets.
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Sequence similaritiesBelongs 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. -
DomainThe 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.
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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 localizationCytoplasm. 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: 785911 Cow
- Entrez Gene: 6093 Human
- Entrez Gene: 19877 Mouse
- Entrez Gene: 81762 Rat
- Omim: 601702 Human
- SwissProt: Q8MIT6 Cow
- SwissProt: Q13464 Human
- SwissProt: P70335 Mouse
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 [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171-239961-ab45171wb.jpg)
Western blot - Anti-ROCK1 antibody [EP786Y] (ab45171)All lanes : Anti-ROCK1 antibody [EP786Y] (ab45171) at 1/5000 dilution (purified)
Lane 1 : HeLa cell lysate - treated with Calyculin A
Lane 2 : HeLa cell lysate - treated with Camptothecin
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : Ramos cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 158 kDa
Observed band size: 158 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171-239964-ab45171ihcp.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EP786Y] (ab45171)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling ROCK1 with purified ab45171 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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![Immunocytochemistry/ Immunofluorescence - Anti-ROCK1 antibody [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171-239965-ab45171iccif.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-ROCK1 antibody [EP786Y] (ab45171)Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ROCK1 with purified ab45171 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150078, an Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/50) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).
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![Flow Cytometry - Anti-ROCK1 antibody [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171-4-ab45171FC.jpg)
Flow Cytometry - Anti-ROCK1 antibody [EP786Y] (ab45171)Overlay histogram showing HeLa cells stained with unpurified ab45171 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45171, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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![Immunoprecipitation - Anti-ROCK1 antibody [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171-239966-ab45171ip.jpg)
Immunoprecipitation - Anti-ROCK1 antibody [EP786Y] (ab45171)ab45171 (purified) at 1/40 immunoprecipitating ROCK1 in HeLa cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Western blot - Anti-ROCK1 antibody [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171-239962-ab45171wb2.jpg)
Western blot - Anti-ROCK1 antibody [EP786Y] (ab45171)All lanes : Anti-ROCK1 antibody [EP786Y] (ab45171) at 1/5000 dilution (purified)
Lane 1 : PC-12 cell lysate
Lane 2 : RAW264.7 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 158 kDa
Observed band size: 158 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171-216496-ab45171ihcp.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EP786Y] (ab45171)This image is courtesy of an anonymous AbreviewUnpurified ab45171 staining ROCK1 in mouse embyro 18dpc tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 2 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/100 in 1% BSA + 1% FBS in TBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EP786Y] (ab45171)](https://www.abcam.com/ps/products/45/ab45171/Images/ab45171_3.bmp)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ROCK1 antibody [EP786Y] (ab45171)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling ROCK1 with unpurified ab45171.
References
This product has been referenced in:
- Wang Y et al. Adipokine RBP4 drives ovarian cancer cell migration. J Ovarian Res 11:29 (2018). Read more (PubMed: 29642915) »
- Wang X et al. Efficacy of extracts of Celastrus orbiculatus in suppressing migration and invasion by inhibiting the EZH2/ROCK1 signaling pathway in human nasopharyngeal carcinoma. Oncol Lett 15:6695-6700 (2018). Read more (PubMed: 29725411) »
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