CaMK2delta Human CAMK Kinase Enzymatic Radiometric LeadHunter Assay [10 μM ATP] - TW

Item: 168040

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OVERVIEW

DOCUMENTS/REFERENCES

Biological Information

Background Information:

Calcium/calmodulin dependent protein kinase II (CaMKII) is a serine/threonine kinase which is encoded as over two dozen variants generated by alternative mRNA splicing of 4 genes. CaMKII phosphorylate target proteins in a calcium calmodulin dependent manner and play critical for gene transcription, ion channel activity, cytoskeletal function and carbohydrate metabolism. CaMKII has been implicated in memory and cell growth control as well as in the disorders of cancer, cystic fibrosis, Alzheimerís disease and epilepsy. The overexpression of a specific constitutively active CAMK2D (CaMKIId) in PC-12, P19 and NIH/3T3 cells induces neurite outgrowth.

Family:

Kinase

Sub Family:

CAMK

Class:

Protein Ser/Thr

Protein Name:

CaMK2delta

Uniprot Number:

Q13557

Protein Aliases:

CaMK-II subunit delta

Gene Name:

CAMK2D

Gene ID:

817

Gene Aliases:

CAMKD

Accession Number:

NM_001321566

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 CAMK2D expressed in insect cells is used. Test compound and/or vehicle is preincubated with 0.2 µg/ml enzyme‡ in modified MOPS buffer pH 7.2 for 15 minutes at 37°C. The reaction is initiated by addition of 50 µM autocamtide-2, 10 µM ATP and 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]Autocamtide-2 formed. Compounds are screened at 10 µM.

Substrate:

50 µM Autocamtide-2

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

98053103

Name

K-252a

Measurement

0.0022μM - IC50

PubChem ID

122130844

Name

Staurosporine

Measurement

0.00047μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Adverse / Beneficial Action:

Inhibition of CAMK2D may impair memory and learning.

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei