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In‐gel detection of urease activity by nitroprusside–thiol reaction
Author(s) -
Sharma Vineet,
Chaudhary Rajesh,
Khurana J. M.,
Muralidhar K.
Publication year - 2007
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.1012
Subject(s) - urease , chemistry , urea , chromatography , polyacrylamide , incubation , staining , biochemistry , polyacrylamide gel electrophoresis , thiol , substrate (aquarium) , enzyme , polymer chemistry , medicine , pathology , geology , oceanography
Abstract A new staining method for urease activity in non‐denaturing polyacrylamide gels is described. The increase in local pH of the gel, resulting from ureolytic activity of urease, causes a purple red coloured band after incubation of the polyacrylamide gel with urea. Staining of urease activity using this method is very specific for catalytically active urease even in crude preparations. Detection of urease activity by this method is rapid, simple and economical. The described method is also more sensitive than existing methods of urease staining. A minimum of 0.25 mU of urease activity can be detected after 5 min of incubation with the substrate. The method has been used to demonstrate the presence of different charge isoforms of urease in a member of the plant family Cucurbitaceae. Copyright © 2007 John Wiley & Sons, Ltd.