AAK1 Human NAK Kinase Enzymatic Radiometric [Km ATP] KinaseProfiler LeadHunter Assay - FR

Procedure Summary:

AAK1(h) is incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 250 µM KEEQSQITSQVTGQIGWR, 10 mM Magnesium acetate 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 120 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the stopped reaction is 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 KEEQSQITSQVTGQIGWR

Tracer:

33P

ATP Concentration:

[ATP]= 45µM (Km= 40µM )

Incubation:

120 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