Biological Information

Background Information:

c-Jun N-terminal kinases (JNKs), a stress-activated protein kinase (SAPK), is involved in cellular responses to environmental stresses. JNK kinases can be activated by stimuli such as UV light, radiation, protein synthesis inhibitors, ceramide, DNA-damaging drugs, chemopreventive drugs, TNF-a, and interleukin 1. JNK binds to an NH2-terminal region of ATF2 and c-Jun and phosphorylates two sites within the activation domain of each transcription factor. Three JNK genes (Jnk1, -2 and -3) have been identified in humans; however, splice variants result in a total of 10 isoforms. Therapeutic inhibition of JNK may provide clinical benefit in disease as diverse as immune inflammatory (joint, bowel, pulmonary), ischemic injury (brain, cardiac) and neurodegenerative (Alzheimer’s, Parkinson’s) diseases.

Family:

Kinase

Sub Family:

CMGC

Class:

Protein Ser/Thr

Protein Name:

JNK1

Uniprot Number:

P45983

Protein Aliases:

JUN N-terminal kinase

Gene Name:

MAPK8

Gene ID:

5599

Gene Aliases:

JNK|JNK1|SAPK1|PRKM8|SAPK1C

Organism:

Human

Construct Details:

Full length recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Radiometric

Measured Response:

Scintillation

Testing Information

Procedure Summary:

JNK1a1 (h) is incubated with 50 mM Tris pH 7.5, 0.1 mM EGTA, 0.1% 6 -mercaptoethanol, 3 µM ATF2, 10 mM MgAcetate and [gamma-33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg/ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting.

Substrate:

3 uM ATF2

Tracer:

33P

ATP Concentration:

[ATP]= 10µM (Km= 41µM )

Incubation:

40 min at Room temperature

Control Inhibitor:

K-252a. Nocardiopsis sp

Test Sample Requirements:

Please provide compounds dissolved in 100% DMSO (PBS or HBSS prohibited).
Stock solution should be ≥50× the final assay concentration (a 10 mM stock is also acceptable).
Required volume:
– <60 kinase assays: 120 µL of 50× stock
– ≥60 assays: 420 µL of 50× stock
If you supply 10 mM stocks or powder (pre-weighed), we will apply the appropriate conversion.
For batches of ~50 compounds supplied as solids, the TAT will be extended.

Minimum Order Quantity:

2

Turnaround Time

Standard:

6 Business Days

Service Scheduling Note:

Assay is run on a weekly schedule beginning on Friday.  Compounds and purchase order must be received on or before 13:00 CET Thursday for the order to be included on the service run that begins on Friday of the same week.

Reference Compound Data

PubChem ID

490561

Name

K-252a, Nocardiopsis sp.

Measurement

145.76nM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

KP - France - Celle L'Evescault

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Item: 14-327KP