analyzed by liquid scintillation by putting them into counting
vials with Ultima Gold. In this case the filter paper will
“quench” the samples (~20%), and therefore the 10μl of the
SA must also be spotted on a piece of filter paper to quench
these accordingly.- Adding 1μCi^33 P-ATP to each sample (0.1μlofa10μCi/μl
stock) (see Note 12) equals 2,220,000 DPM
(1μCi¼37,000 Bq¼2,220,000 DPM). The kinase reaction
buffer was diluted 67 times in water (15μl plus 985μl of water)
(seestep 6of the Subheading3.2), and only 10μl was added to
the counting vial while 30μl was added to the samples, giving
an extra three times dilution making the total dilution
200 times. The counting values obtained measuring the SA
should therefore be approximately 11,100 CPM or DPM
(2,220,000/200¼11,100).
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