Recombinant Anti-ROCK2 antibody [EPR7141(B)] (ab125025)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7141(B)] to ROCK2
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ROCK2 antibody [EPR7141(B)]
See all ROCK2 primary antibodies -
Description
Rabbit monoclonal [EPR7141(B)] to ROCK2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human ROCK2 (C terminal). The exact sequence is proprietary.
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Positive control
- HepG2, A549, HeLa, HEK293, L6, RAW264.7 and A673 cell lysates; Human kidney tissue, Mouse and Rat brain tissue 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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 8.70 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.14% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7141(B) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Assay kits
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab125025 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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WB | (3) |
1/10000 - 1/50000. Detects a band of approximately 161 kDa (predicted molecular weight: 161 kDa).
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ICC/IF |
1/100 - 1/250.
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Notes |
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WB
1/10000 - 1/50000. Detects a band of approximately 161 kDa (predicted molecular weight: 161 kDa). |
ICC/IF
1/100 - 1/250. |
Target
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Function
Regulates the assembly of the actin cytoskeleton. Promotes formation of stress fibers and of focal adhesion complexes. Plays a role in smooth muscle contraction. -
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. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Cytoplasm. Cell membrane. Cytoplasmic, and associated with actin microfilaments and the plasma membrane. - Information by UniProt
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Database links
- Entrez Gene: 9475 Human
- Entrez Gene: 19878 Mouse
- Entrez Gene: 25537 Rat
- Omim: 604002 Human
- SwissProt: O75116 Human
- SwissProt: P70336 Mouse
- SwissProt: Q62868 Rat
- Unigene: 58617 Human
see all -
Alternative names
- coiled-coil-containing protein kinase 2 antibody
- KIAA0619 antibody
- p164 ROCK 2 antibody
see all
Images
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ROCK2 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A10 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab125025 observed at 165 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab125025 was shown to specifically react with ROCK2 when ROCK2 knockout samples were used. Wild-type and ROCK2 knockout samples were subjected to SDS-PAGE. ab125025 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 h at room temperature before imaging. -
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling ROCK2 with purified ab125025 at 1/250. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291 anti-Tubulin (mouse mAb) primary and ab150120 (AlexaFluor®594 goat anti-mouse) secondary both at 1/1000 dilution. Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody and ab150120 (anti-mouse) secondary antibody were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by ab150077 (anti-rabbit secondary antibody).
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All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ROCK2 knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 161 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab125025 observed at 175 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab125025 was shown to react with ROCK2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265679 (knockout cell lysate ab257643) was used. Wild-type HeLa and ROCK2 knockout 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. ab125025 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 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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All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/50000 dilution
Lane 1 : HeLa whole cell lysate
Lane 2 : HEK293 whole cell lysate
Lane 3 : Mouse brain tissue lysate
Lane 4 : Rat brain tissue lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 161 kDa
Observed band size: 161 kDa
Blocking and Diluting buffer 5% NFDM/TBST -
All lanes : Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution (unpurified)
Lane 1 : Hela cell lysate
Lane 2 : L6 cell lysate
Lane 3 : A673 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 161 kDa -
Anti-ROCK2 antibody [EPR7141(B)] (ab125025) at 1/10000 dilution + RAW264.7 whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 161 kDa
Observed band size: 161 kDa
Blocking and Diluting buffer 5% NFDM/TBST
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (39)
ab125025 has been referenced in 39 publications.
- Li C et al. Rho kinase inhibition ameliorates vascular remodeling and blood pressure elevations in a rat model of apatinib-induced hypertension. J Hypertens 40:675-684 (2022). PubMed: 34862331
- Chen R et al. CID16020046, a GPR55 antagonist, attenuates sepsis‑induced acute kidney injury. Mol Med Rep 25:N/A (2022). PubMed: 35244189
- Tonon F et al. 5-Azacytidine Downregulates the Proliferation and Migration of Hepatocellular Carcinoma Cells In Vitro and In Vivo by Targeting miR-139-5p/ROCK2 Pathway. Cancers (Basel) 14:N/A (2022). PubMed: 35406401
- Feng XF et al. Magnetic Resonance Imaging Investigation of Neuroplasticity After Ischemic Stroke in Tetramethylpyrazine-Treated Rats. Front Pharmacol 13:851746 (2022). PubMed: 35559236
- Zhao X et al. Rock1 is a novel host dependency factor of human enterovirus A71: Implication as a drug target. J Med Virol 94:5415-5424 (2022). PubMed: 35791459