Garlic Increases Antioxidant Levels in Diabetic and Hypertensive Rats Determined by a Modified Peroxidase Method

Table 1

Protocol for the antioxidant assay.

Solution	Blank^a (μL)	Sample (μL)
PBS (5 mM, pH 7.4, 138 mM NaCl)	1000 – (105 + x)	1000 – (115 + x)
MetMb (as determined above)	𝑥^b	x
Sample (serum or antioxidant)	—	10
ABTS (5 mM)	30	30
Equilibrate for 3 min at 30°C
H2O2 (10 mM)	75	75
Incubate for 6 min at 30°C
Read absorbance at 734 nm after exactly 15 s of incubation period completion^c

^a Read against PBS buffer.
^b MeMb concentration is determined experimentally and the required volume (x) is added to obtain 2.5 𝜇M final concentration in the assay. The volume of PBS buffer is adjusted accordingly.
^c Only six samples can be run at a time to fit the timing protocol.