Recombinant Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free (ab219587)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1 - BSA and Azide free
- Suitable for: IHC-Fr, WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Cow, Human
Related conjugates and formulations
Overview
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Product name
Anti-ROCK2 + ROCK1 antibody [EP786Y] - BSA and Azide free -
Description
Rabbit monoclonal [EP786Y] to ROCK2 + ROCK1 - BSA and Azide free -
Host species
Rabbit -
Specificity
This 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 and has been shown to bind recombinant human ROCK2, please see western blot images below.
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Tested applications
Suitable for: IHC-Fr, WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Cow, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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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 (intra): HeLa cells. IP: HeLa cell lysate.
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General notes
ab219587 is the carrier-free version of ab45171.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP786Y -
Isotype
IgG -
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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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab219587 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 |
Use at an assay dependent concentration. PubMed: 19295659
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 158 kDa.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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IHC-Fr
Use at an assay dependent concentration. PubMed: 19295659 |
WB
Use at an assay dependent concentration. Predicted molecular weight: 158 kDa. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Cellular localization
ROCK2: Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane. ROCK1: 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. -
Database links
- Entrez Gene: 282041 Cow
- Entrez Gene: 785911 Cow
- Entrez Gene: 6093 Human
- Entrez Gene: 9475 Human
- Entrez Gene: 19877 Mouse
- Entrez Gene: 19878 Mouse
- Entrez Gene: 25537 Rat
- Entrez Gene: 81762 Rat
see all
Images
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ab45171 (purified) at 1/40 immunoprecipitating ROCK2 + 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling ROCK2 + 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human adenocarcinoma of the colon tissue labelling ROCK2 + 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).
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All lanes : Anti-ROCK2 + ROCK1 antibody [EP786Y] (ab45171) at 1/1000 dilution
Lane 1 : Recombinant Human ROCK1 protein (aa 1114 to 1354) (30 kDa)
Lane 2 : Recombinant Human ROCK2 protein (aa 1132 to 1388) (30 kDa)
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 158 kDa
Observed band size: 30 kDa why is the actual band size different from the predicted?This data was developed using ab45171, the same antibody clone in a different buffer formulation.
Loading control: Anti-6X His tag® antibody [EPR20547] (ab213204)
Blocking buffer and concentration: 5% NFDM/TBST
Exposure Times:
Lane 1: 3 seconds
Lane 2: 7 seconds
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling ROCK2 + ROCK1 with unpurified ab45171.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45171).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (24)
ab219587 has been referenced in 24 publications.
- Cheng C et al. MicroRNA-145-5p inhibits hypoxia/reoxygenation-induced apoptosis in H9c2 cardiomyocytes by targeting ROCK1. Exp Ther Med 22:796 (2021). PubMed: 34093752
- Buonpane C et al. ROCK1 inhibitor stabilizes E-cadherin and improves barrier function in experimental necrotizing enterocolitis. Am J Physiol Gastrointest Liver Physiol 318:G781-G792 (2020). PubMed: 32090605
- Lin LL et al. PAI-1-Dependent Inactivation of SMAD4-Modulated Junction and Adhesion Complex in Obese Endometrial Cancer. Cell Rep 33:108253 (2020). PubMed: 33053339
- Huang ZX et al. RhoA/ROCK pathway mediates the effect of oestrogen on regulating epithelial-mesenchymal transition and proliferation in endometriosis. J Cell Mol Med 24:10693-10704 (2020). PubMed: 32725958
- Gentry EG et al. Rho Kinase Inhibition as a Therapeutic for Progressive Supranuclear Palsy and Corticobasal Degeneration. J Neurosci 36:1316-23 (2016). PubMed: 26818518