A simple, reliable and accurate method for the measurement of D-hexuronic acids (specifically D-glucuronic acid and D-galacturonic acid) in plant extracts, culture media/supernatants and other materials.
Suitable for manual, auto-analyser and microplate formats.
INTRODUCTION:
D-glucuronic acid and D-galacturonic acid are naturally occurring
hexuronic acids present in glycosaminoglucans, glucuronide conjugates
in mammals and in plant cell wall polysaccharides.
D-Glucuronic acid exists as a component of glycosaminoglucans such
as hyaluronan, heparin and chondroitin sulphate present in mammalian
connective tissue such as cartilage. In the synthesis of glucuronide
conjugates in mammals glucuronic acid is conjugated to xenobiotic
compounds, a biosynthesis reaction known as glucuronidation,
catalysed by the enzyme UDP-glucuronyltransferase and considered a
detoxifying process.
Both D-glucuronic acid and D-galacturonic acid are major components
of plant cell wall polysaccharides. D-glucuronic acid is a component
of arabinoxylan, which consists of a β-(1,4)-linked xylan backbone
substituted with α-(1-2/3)-linked L-arabinofuranose and α-(1-2)-
linked 4-O-methylglucuronic acid. D-Galacturonic acid is the major
component of pectin comprising the α-(1,4)-linked galacturonan
backbone of homogalacturonan and rhamnogalacturonan II, and is
present within the repeating disaccharide unit [,4)-α-D-GalpA-(1,2)-α-
L-Rhap-(1,] of rhamnogalacturonan I.
This kit (K-URONIC) is suitable for the specific measurement of
D-hexuronic acids including D-glucuronic and D-galacturonic acid.
KITS:
Kits suitable for performing 100 assays in manual format (or 1000
assays in auto-analyser format or 1000 assays in microplate format)
are available from Megazyme. The kits contain the full assay method
plus:
Bottle 1: Buffer (22 mL, pH 8) plus sodium azide (0.02% w/v)
as a preservative.
Stable for > 2 years at 4°C.
Bottle 2: NAD+ freeze dried powder.
Stable for > 5 years at -20°C.
Bottle 3: Uronate dehydrogenase suspension (2.2 mL).
Stable for > 2 years at 4°C.
Bottle 4: D-Glucuronic acid (5 mL; 0.5 mg/mL)
Stable for > 2 years at 4°C.