Browsing by Subject "Isoperoxidasas"

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    Quantification of lupin peroxidase isoenzymes by densitometry
    (Murcia: Universidad de Murcia, Servicio de Publicaciones, 1987) Ros Barceló, Alfonso; Facultad de Biología
    The formation of benzidine-brown end product by lupin peroxidase isoenzymes, separated by gel isoelectric focusing, is used in order to quantify isoenzyme activities on the same gel. Incubation times of 3 h, H2O2 concentration of 1 mM, ben~ -as saturated solution, and pH 5.0, are the best assay conditions. Under these conditions, isoperoxidase activity can be quantified as formation (and/or acéumulation) of the benzidine-brown end product in the steady-state ofthe reaction. The product can be measured as the peak area recorded· by scanning densitometry at 460 nm. Linear relationship between the amount of total peroxidase activity loaded on the top of the gel, and the area of each isoperoxidase peak resolved by gel isoelectric focusing support the validity of this densitometric method for the quantification of lupin peroxidase isoenzymes.