PRAK Human CAMK Kinase Enzymatic Radiometric LeadHunter Assay [10 μM ATP] - TW

Item: 171380

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OVERVIEW

DOCUMENTS/REFERENCES

Biological Information

Background Information:

Protein Serine/Threonine Kinase MAPKAPK5 (mitogen-activated protein kinase activated protein kinase 5), also known as p38-regulated/activated protein kinase (PRAK), functions downstream of p38a and p38b in mediating signaling of the p38 pathway. This kinase can be activated in response to cellular stress and proinflammatory cytokines through its phosphorylation by MAP kinases including MAPK1/ERK, MAPK14/p38-alpha, and MAPK11/p38-beta. MAPKAPK5 is localized predominantly in the cytoplasm and is constantly shuttled between the cytoplasm and the nucleus. MAPKAPK5 may play a role in balancing other MAPK pathways because overactivation of MAPKAPK5 can inhibit Ras mediated cell proliferation and gene activation. MAPKAPK5 inhibtion may facilitate cell differentiation, inflammation and cell proliferation.

Family:

Kinase

Sub Family:

CAMK

Class:

Protein Ser/Thr

Protein Name:

PRAK

Uniprot Number:

Q8IW41

Protein Aliases:

Gene Name:

MAPKAPK5

Gene ID:

8550

Gene Aliases:

PRAK

Accession Number:

NM_139078

Organism:

Human

Construct Details:

Full Length, 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 PRAK expressed in insect Sf21 cells is used. Test compound is preincubated with 0.44 U/ml enzyme‡ in modified MOPS buffer pH 7.2 for 15 minutes at 37°C. The reaction is initiated by addition of 30 μM PRAK substrate, 10 μM ATP containing 0.25 μCi[γ32P]ATP for another 30 minute incubation period and terminated by further addition of 3% H3PO4. An aliquot is removed and counted to determine the amount of [32P]PRAK substrate. Compounds are screened at 10 μM.

Substrate:

30 µM PRAK substrate peptide

ATP Concentration:

10 uM

Incubation:

30 min at 37°C

Control Inhibitor:

Staurosporine

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:

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

176155

Name

SB 203580

Measurement

>10μM - IC50

PubChem ID

5281847

Name

Rottlerin

Measurement

4.6μM - IC50

PubChem ID

122130844

Name

Staurosporine

Measurement

0.29μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei