Detecting calcium-binding proteins?

Fergus Doherty mbxfd at unicorn.nott.ac.uk
Thu Mar 17 12:02:18 EST 1994


Hello,
Does anyone know of a simple way to detect calcium binding proteins?  I've
come across reference to the dye Stains-all which binds to calcium binding
proteins and is displaced by calcium.  Stains-all stains all proteins red
apparently, but calcium binding proteins blue.  An article in 1988
(Urban,Ancel, Virmaux, Biochem Soc Trans, 16, 299) suggests that they
detected a calcium binding protein by staining an SDS-PAGE gel (after some
renaturation?), but there are no details.  Anyone come across this?  Would
it stain a nitrocellulose transfer?  I know you can look at spectral shifts
in Stains-all in a spectrophotometer with the test protein but I might be
pushed for enough of it, and its not pure.  I have a limited amount of this
protein (few mgs, contaminated with one other major protein) I want to test
for calcium binding, any suggestions. 

Fergus Doherty,
dept. Biochemistry,
University Medical School,
Queen's Medical Centre,
Nottingham NG7 2UH
Tel: 602 709366  FAX 602 422225 Internet: mbxfd at unicorn.nott.ac.uk



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