Protein phos assays, TCA and dot blot manifolds

Michael Dahl dahl at
Wed Nov 18 09:16:45 EST 1992

pcovello at writes:

>Speaking of protein phosphorylation assays, I have a 
>question along similar lines. I am looking for a very
>rapid method for TCA precipitation/scintillation 
>counting of nucleic acids. I have read that a vacuum
>manifold can be useful for washing filters and I 
>notice that the "Minifold" on p. 20 of the 1992
>Schleicher and Schuell catalogue is recommended
>for this purpose. Does anyone have any experience
>using a vacuum manifold for this purpose. Can anyone
>recommend a protocol for its use?

>Pat Covello
>Dept. Biochemistry
>Dalhousie University
>Halifax, NS

Dear Pat,

I do not have the catalogue from Schleicher and Schuell-anyway, may be
I can help you. I used a very rapid method for the DegS phosphorylation
of Bacillus sublitis for the determination of its Km for ATP. The Method is described in our paper: Dahl et al. 1991, J.Bacteriol. p 2539-2547. You need
nitrocellulose filters (0.05 micrometers of pore size). 
The method as described is really rapid and simple to handle.
If you have more questions feel free ad contact me.
dahl at

Michael Dahl
Universitaet Freiburg (Biologie III)
Schaenzlestrasse 1

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