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A novel colorimetric assay and activity staining for cytokinin oxidase based on the copper amine oxidase property of the enzyme
Author(s) -
Kulkarni Mahesh J.,
Theerthaprasad D.,
Sudharshana L.,
Prasad T. G.,
Sashidhar V. R.
Publication year - 2001
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.581
Subject(s) - chemistry , amine oxidase , copper , oxidase test , enzyme , amine gas treating , colorimetry , formazan , staining , chromatography , biochemistry , organic chemistry , medicine , pathology
A rapid and sensitive colorimetric assay for cytokinin oxidase (CKO) was developed based on the copper amine oxidase property of the enzyme from cucumber cotyledons. The assay involved oxidation of isopentenyl adenine by CKO in the presence of phenazine methosulphate as an electron acceptor, resulting in reduction of nitroblue tetrazolium to formazon. The formation of formazon was linear with time for at least 20 min and could be followed spectrophotometrically at A 490 . CKO could also be specifically stained on polyacrylamide gels using the same principle. Copyright © 2001 John Wiley & Sons, Ltd.