RabMAb

Anti-ROCK1抗体[EP786Y] (ab45171)

概述

  • 产品名称Anti-ROCK1抗体[EP786Y]
    参阅全部 ROCK1 一抗
  • 描述
    兔单克隆抗体[EP786Y] to ROCK1
  • 特异性This antibody recognizes both the cleaved C-terminus of ROCK 1 (30 kDa) and full length protein (158 kDa).
  • 经测试应用适用于: IHC-Fr, WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Cow, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human ROCK1 aa 1100-1200 (C terminal). The exact sequence is proprietary.
    Database link: Q13464

  • 阳性对照
    • 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.
  • 常规说明

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    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.

性能

应用

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.

应用 Ab评论 说明
IHC-Fr 1/100. PubMed: 19295659
WB 1/2000 - 1/10000. Predicted molecular weight: 158 kDa.

For unpurified use at 1/500.

IHC-P 1/50 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/50 - 1/500.

 

IP 1/40 - 1/50.

 

Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

靶标

  • 功能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.
  • 组织特异性Detected in blood platelets.
  • 序列相似性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.
  • 结构域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.
  • 翻译后修饰Autophosphorylated on serine and threonine residues.
    Cleaved by caspase-3 during apoptosis. This leads to constitutive activation of the kinase and membrane blebbing.
  • 细胞定位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
  • 数据库链接
  • 别名
    • 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

Anti-ROCK1 antibody [EP786Y] 图像

  • 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
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • 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.

  • 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).

  • 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.

  • 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.

  • 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
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Unpurified 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.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling ROCK1 with unpurified ab45171.

Anti-ROCK1 antibody [EP786Y] (ab45171)参考文献

This product has been referenced in:
  • Gentry EG  et al. Rho Kinase Inhibition as a Therapeutic for Progressive Supranuclear Palsy and Corticobasal Degeneration. J Neurosci 36:1316-23 (2016). Read more (PubMed: 26818518) »
  • Zhang GM  et al. MicroRNA-126 inhibits tumor cell invasion and metastasis by downregulating ROCK1 in renal cell carcinoma. Mol Med Rep 13:5029-2036 (2016). WB ; Human . Read more (PubMed: 27108693) »

See all 20 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 Nov 04 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step FBS / BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9
Sample Human Tissue sections (Infantile fibromatosis)
Specification Infantile fibromatosis
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 04 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step FBS / BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9
Sample Mouse Tissue sections (Mouse embryo 18dpc)
Specification Mouse embryo 18dpc
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jan 15 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (MEF)
Loading amount 30 µg
Specification MEF
Treatment TGF b1 o/n 5 ug/ul
Gel Running Conditions Reduced Denaturing (7.5)
Blocking step (agent) for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Feb 27 2013

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...

Read More

Thank you once again for your message. Thank you also for your patience waiting for a response, I wanted to ensure I had time to review the whole case again fully before contacting you as it has been a while since we last corresponded.

I am ...

Read More

Thank you for your message. I am sorry to hear that the alternative free of charge replacement has also not worked.

It is regrettable both the antibodies have not provided satisfactory results. I would like to reassure you that ab97592 is te...

Read More

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, a...

Read More

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 h...

Read More

Thank you for confirming these details and for your cooperation.

I am sorry that ab45171 has not been working for you and for the inconvenience this has caused. As requested, I have issued an alternative free of charge replacement of 97592 w...

Read More

1-10 of 17 Abreviews or Q&A

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