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PMID: 2472757 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Selective silver staining of urease activity in polyacrylamide gels.

Analytical biochemistry ·Vol. 177 ·No. 1 ·1989-02-15 ·Pages 37-40

Martín de Llano JJ, García-Segura JM, Gavilanes JG

Abstract

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.

MeSH Terms
Densitometry Electrophoresis, Polyacrylamide Gel Polymers Silver Staining and Labeling/methods Urease/analysis
Chemicals
Polymers Silver Urease
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Martín de Llano J J
Departmento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad Complutense, Madrid, Spain.
García-Segura J M
Gavilanes J G
Article Info
Journal
Analytical biochemistry
Abbr.
Anal Biochem
ISSN
0003-2697
Published
1989-02-15
Pages
37-40
Language
English
Region
United States
NLM ID
0370535
Subset
IM
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