The ABEL 60 series are chemiluminescent test kits for quantifying low levels of superoxide produced as a by product in the production of uric acid from the enzyme-catalysed oxidation of xanthine with xanthine oxidase.The assay can be used to quantify the superoxide produced by cells as well as for assessing the antioxidant capacity of therapeutic reagents and ingredients in foods, nutraceuticals and cosmetics. The activity of superoxide dismutases and mimetics of this enzyme can be quantified as well as inhibitors of xanthine oxidase.

CONTENTS

Kit components sufficient for 100 x 200μL tests

A. 1 x bottles 50μg Pholasin (reconstitute to 5 mL)
B. 1 x 50mL Reconstitution & Assay Buffer for xanthine oxidase.
C. 1 x 10mL Reconstitution Buffer for xanthine
D. 1 x bottles of xanthine (reconstitute with 2.5mL buffer to obtain 16mM)
E. 1 x bottles xanthine oxidase: 51.25mU
F. 1 x bottles superoxide dismutase: 125 U
G. 1 x 96 well white microplates

The ABEL 61 series are chemiluminescent test kits for quantifying superoxide(anion radical 02·-) produced as a by product of the production of uric acid from the enzyme reaction of xanthine with xanthine oxidaseand for use as an antioxidant test and for measuring the activity of superoxide dismutase (SOD) The assay can be used to quantify the superoxide producedby cells as well as assessing the antioxidant capacity of therapeutic reagents and ingredients in foods, nutraceuticals and cosmetics.One unit of xanthine oxidase catalyses the oxidation of 1μmol xanthine to uric acid with theconcomitant production of 2μmol superoxide per minute at 250C with the rate of reaction doubling for approximatelyevery 100C increase in temperature Pholasin, an ultrasensitve chemiluminescent detector of superoxide can detect concentrations in this assay as low as 50fmol per minute, which is in the order produced by small numbers of cells. Higher amounts of superoxide can also be generated for use in antioxidant and other assays.

This assay has many advantages over the non-specific indirect cytochrome C method. The activity of superoxide dismutases and mimetics of this enzyme can be quantified very easily. As SOD will compete with Pholasin? for any superoxide produced in x/xo system less light will be emitted in the presence of SOD. From a set of SOD standards the amount of SOD or a mimetic of SOD in a sample to be tested can be determined by the amount of light emitted in the presence of Pholasin.