effect of infection of the filarial parasite brugia malayi - Pondicherry ...

4.1.1.6 Glucose-6-phosphate dehydrogenase
modifications.
This enzyme was assayed by the method of Z i et a/. (1958) with some
Glucose-&phosphate dehydrogenase is assayed by measuring the increase in
absorbance, which occurred at 340 nm when NADP is reduced to NADPH. The reduction
takes place when two electrons are transferred fkom glucose-6-phosphate to NADP in the
reaction catalysed by glucose-&phosphate dehydrogenase.
Reagents
I. NADP : 0.2 mM
2. Tris-HCI buffer : 0. l M, pH 8.0
3. Magnesium chloride : 0.1 M
4. Glucose-6-phosphate : 6 mM
To 0.1 ml of NADP, buffer and MgC12, 0.5 ml of water and 0.1 ml of the
homogenate was added. After 10 minutes, the reaction was initiated by the addition of 0.1 ml
of glucose-6-phosphate solution. The increase in optical density was measured at 340 nm at
2S°C. The activity of the enzyme was expressed in terms of units/mg/protein where one unit
cornponds to the amount of the enzyme required to bring about a change in optical density
of 0.01 /minute.
4.1.1.7 Reduced glutathione
Reduced glutathione was estimated by the method of Moron eta/. (1979)
This method is based on the development of yellow colour when 5,5'dithio-bis-2-
n~trobic acid (DTNB) is added to the compounds containing sulphydryl pups.