Sensitive alkaline phosphatase assay

Ivan Torshin TIY at poltorak.chem.msu.su
Sat Jun 20 11:11:53 EST 1998


>
>but the alkaline phosphatase activity is too low to be useful.
>

Dear Dudley Page,

Could you describe the experiment in more detail ?
For example, what APAse do you use (source, another data) ?
What is the expression system (E.coli) ? Do you experiment
with pH, temperature (APAse is periplasmic protein, therefore, it could be
easily regulated by changing 'outside' pH. E.coli Apase is relatively
thermostable- the molecule begins to dissociate into subunits with the activity
loss at 70-80 C) ? Moreover, E.coli APase activity strongly depends
on the MG and Zn ion concentrations -the ions are involved in active site
and molecule stabilization)

Some of that may help,

Regards,
Ivan






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