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_172115

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:

CaMKIId (h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.5 mM CaCl2, 16 ug/mL calmodulin, 250 µM KKLNRTLSFAEPG, 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:

250 µM KKLNRTLSFAEPG

Tracer:

33P

ATP Concentration:

[ATP]= 10µM (Km= 10.11µM )

Incubation:

40 min at Room temperature

Control Inhibitor:

Staurosporine

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

122130844

Name

Staurosporine

Measurement

0.23nM - 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:

KP - France - Celle L'Evescault

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Item: 14-723KP