Biological Information

Background Information:

Rho-associated, coiled-coil containing protein kinase (ROCK), including the ROCK1 and ROCK2 isoforms, is a protein kinase involved in signaling from Rho to actin cytoskeleton. PKA and the two ROCKs have been reported to have opposite effects on neurite formation. More studies are needed to provide significant insights into the role of ROCK in several pathological processes. ROCK inhibitors have been shown the benefit for treatment of various cancers, inflammation, coronary artery spasms and hypertension.

Family:

Kinase

Sub Family:

AGC

Class:

Protein Ser/Thr

Protein Name:

ROCK2

Uniprot Number:

O75116

Protein Aliases:

Rho kinase 2|Rho-associated, coiled-coil-containing protein kinase 2|Rho-associated, coiled-coil-containing protein kinase II|ROCK-II|p164 ROCK-2

Gene Name:

ROCK2

Gene ID:

9475

Accession Number:

NM_001321643

Organism:

Human

Construct Details:

Fragment (11-552), Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Radiometry

Measured Response:

Scintillation

Testing Information

Procedure Summary:

Human recombinant protein kinase ROCK2 expressed in Sf21 insect cells is used. Test compound and/or vehicle is preincubated with 81 mU/ml enzyme‡ for 15 minutes at 37°C in modified MOPS buffer pH 7.2. The reaction is initiated by addition of 15 μM long S6 kinase substrate, 10 μM ATP and 0.05 μCi [γ³²P]ATP for another 30 minutes incubation period and terminated by further addition of 3% H3PO4. An aliquot is removed and counted to determine the amount of [32P]Long S6 kinase substrate formed. Compounds are screened at 10 μM.

Substrate:

15 µM Long S6 kinase substrate peptide

ATP Concentration:

10 uM

Incubation:

30 min at 37°C

Control Inhibitor:

Rho-Kinase inhibitor

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

10

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

448042

Name

Y-27632

Measurement

0.2μM - IC50

PubChem ID

122130844

Name

Staurosporine

Measurement

0.0005μM - IC50

PubChem ID

Name

Rho-Kinase inhibitor

Measurement

0.014μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei