Selective silver staining of urease activity in polyacrylamide gels.
A selective method for staining urease activity bands in nondenaturing polyacrylamide gels is described. It is based on the deposition of silver at the urease bands after incubation of gels in the presence of urea and photographic developers. Its highly sensitivity (up to 0.015 enzyme units, corresponding to 5 ng of purified urease) is based on both the silver deposition enhancement methodology and the developers used. The selectivity of the procedure is based on the local pH increase catalytically produced by the enzyme in the presence of urea. The densitometric scan of the enzyme bands gives a linear response at least in the range 0.015-0.300 urease units. This selective staining method is about 2.5 times more sensitive than the standard silver staining of proteins, in terms of detectable urease amount.
Martín de Llano
J J
JJ
Departmento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad Complutense, Madrid, Spain.
García-Segura
J M
JM
Gavilanes
J G
JG
D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D011108:Polymers; D012834:Silver; D013194:Staining and Labeling / Q000379:methods; D014510:Urease / Q000032:analysis*
D011108:Polymers; D012834:Silver; D014510:Urease