Assay of protein kinase using synthetic peptides.

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Dear ABRFers,

I am from Yokohama in japan. Please allow me to send questions.

Now, I do kinase assay using synthetic peptides.
In general protocol, they use TCA for precipitation and phosphoric acid for
washing.
Additionaly, this looks general, I use radio-isotope for labeling and
TOF-MASS exp.

1. Even if you do not use kinase, there is ~5k cpm.Is this nonspecific
binding for peptides?
(In my experience, level of cpm looks the length of peptides.)

2. In case of TOF analysis (w/o RI), control peptides show different
predicted size.
Difference is small(0.1%). I think this is caused by pH or isotope etc.....?
(of course, after kinase reaction, I can get 80Da shift of size.)

Now is the starting periods of new semester, so you all may be
busy....Thanks in advance.

Take

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