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#DBGD-100 QuantiChrom™ â_Glucosidase Assay Kit
Product name : QuantiChrom™ â_Glucosidase Assay Kit
Catalog number : DBGD-100
Supplier name : Bioassays
Data sheet: Ask more or other datasheet now !
About this Product :QuantiChrom™ â_Glucosidase Assay Kit antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of QuantiChrom™ â_Glucosidase Assay Kit.
More Details about
b-GLUCOSIDASE is a glucosidase enzyme which acts upon β1->4 bonds
linking two glucose or glucose-substituted molecules (i.e., the disaccharide
cellobiose). b-Glucosidases are required by organisms (some fungi,
bacteria, termites) for consumption of cellulose. Lysozyme is also a b-
glucosidase and is present in tears to prevent bacterial infection of the eye.
In humans, lower activity of a b-glucosidase isoform (lysosomal glucocerebrosidase)
has been related to Gaucher's disease and Parkinson's
Simple, direct and automation-ready procedures for measuring b-
glucosidase activity are becoming popular in Research and Drug Discovery.
BioAssay Systems' QuantiChromTM b-Glucosidase Assay Kit is designed to
measure b-glucosidase activity directly in biological samples without
pretreatment. The improved method utilizes p-nitrophenyl-b-Dglucopyranoside
that is hydrolyzed specifically by b-glucosidase into a
yellow colored product (maximal absorbance at 405nm). The rate of the
reaction is directly proportional to the enzyme activity.
High sensitivity and wide linear range. Use 20 μL sample. The detection
limit is 2 U/L, linear up to 250 U/L.
Homogeneous and simple procedure. Simple “mix-and-measure”
procedure allows reliable quantitation of b-glucosidase activity within 20
Robust and amenable to HTS. All reagents are compatible with highthroughput
liquid handling instruments.
Direct Assays: b-glucosidase activity in biological samples.
Characterization and Quality Control for b-glucosidase production.
Drug Discovery: high-throughput screen for b-glucosidase modulators.
KIT CONTENTS (100 tests in 96-well plates)
Assay Buffer: 24 mL (pH 7.0)
b-NPG Substrate: 1 mL
Calibrator: 10 mL (equivalent to 250 U/L)
Storage conditions. The kit is shipped at room temperature. Store all
components at -20°C. Shelf life of 6 months after receipt.
Precautions: reagents are for research use only. Normal precautions for
laboratory reagents should be exercised while using the reagents. Please
refer to Material Safety Data Sheet for detailed information.
PROCEDURES. This assay is based on a kinetic reaction. Use of a
multi-channel pipettor is recommended. Addition of Working Reagent to
samples should be quick and mixing should be brief but thorough. Assays
can be executed at room temperature or 37°C.
Reagent preparation: equilibrate reagents to room temperature. The
Working Solution is prepared by mixing for each 96-well assay, 200 μL
Assay Buffer and 8 μL b-NPG substrate (final 1.0 mM). Fresh reconstitution
is recommended, although the Working Solution is stable for at least one
day at room temperature.
Sample preparation: enzyme samples can be in 50 mM phosphate (pH 7.0)
buffer or in any other suitable enzyme buffer. The following chemicals are
known to affect the enzyme activity and should be avoided. SH-containing
reagents (e.g. dithiothreitol, 2-mercaptoethanol, glutathione), Ca2+, Cu2+,
Fe3+/Fe2+, Hg2+, Mg2+, Ni2+, Zn2+, SDS, EDTA and Tris.
Procedure using 96-well plate:
1. Transfer 20 μL distilled water (H2O) to two wells of a clear bottom 96-well
plate. Add 200 μL H2O to one of these wells and 200 μL Calibrator to the
other well (total volume 220 μL).
Transfer 20 μL samples into other wells. Transfer 200 μL Working
Reagent to the sample wells only. The final reaction volume in the
sample wells is 220 μL. Tap plate briefly to mix.
2. Read OD405nm (t = 0), and again after 20 min (t = 20 min) on a plate reader.
4. Calculation: b-glucosidase activity of the sample (U/L) is
OD20 – OD0
ODCALIBRATOR – ODH2O
b-Glucosidase Activity × 250 (U/L)
OD20 and OD0 are OD405nm values of sample at 20 and 0 min,
respectively. ODCALIBRATOR and ODH2O are OD405nm values of Calibrator
and H2O at 20 min.
Unit definition: one unit of enzyme catalyzes the hydrolysis of 1 μmole of
substrate per min at pH 7.0.
MATERIALS REQUIRED, BUT NOT PROVIDED
Pipeting devices and accessories (e.g. multi-channel pipettor). Clear
bottom 96-well plates (e.g. Corning Costar) and plate reader.
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