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:

ROCK1

Uniprot Number:

Q13464

Protein Aliases:

Renal carcinoma antigen NY-REN-35|Rho-associated, coiled-coil-containing protein kinase 1|Rho-associated, coiled-coil-containing protein kinase I|ROCK-I|p160 ROCK-1|p160ROCK

Gene Name:

ROCK1

Gene ID:

6093

Gene Aliases:

p160ROCK

Accession Number:

NM_005406

Organism:

Human

Construct Details:

Fragment (17-535), 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 ROCK1 expressed in insect Sf21 cells is used. Test compound and/or vehicle is preincubated with 0.13 U/ml enzyme‡ in modified MOPS buffer pH 7.2 for 15 minutes at 37°C. The reaction is initiated by addition of 15 μM long S6 kinase substrate, 10 μM ATP and 0.25 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

2.4μM - IC50

PubChem ID

122130844

Name

Staurosporine

Measurement

0.0012μM - IC50

PubChem ID

Name

Rho-Kinase inhibitor

Measurement

0.71μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei