beta-Glucosidase Assay Kit

Catalog No. ABIN1000251

Product Details

Target: Glucosidase, beta

Detection Range: 2-250 U/L

Minimum Detection Limit: 2 U/L

Application: Activity Assay (AcA)

Characteristics: 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 beta-glucosidase activity within 20 minutes.
Robust and amenable to HTS. All reagents are compatible with high- throughput liquid handling instruments.

Components: Assay Buffer: 24 mL (pH 7.0). beta-NPG Substrate: 1 mL. Calibrator: 10 mL (equivalent to 250 U/L).

Material not included: Pipeting devices and accessories (e.g. multi-channel pipettor). Clear bottom 96-well plates (e.g. Corning Costar) and plate reader.

Application Details

Application Notes: Direct Assays: beta-glucosidase activity in biological samples.
Characterization and Quality Control for beta-glucosidase production.
Drug Discovery: high-throughput screen for beta-glucosidase modulators.

Protocol: 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.

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.

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 -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), Ca 2+ , Cu 2+ , Fe 3+ /Fe 2+ , Hg 2+ , Mg 2+ , Ni 2+ , Zn 2+ , SDS, EDTA and Tris.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Mallery, Budendorf, Larsen, Pei, Tong, Holpuch, Larsen, Stoner, Fields, Chan, Ling, Liu: "Effects of human oral mucosal tissue, saliva, and oral microflora on intraoral metabolism and bioactivation of black raspberry anthocyanins." in: Cancer prevention research (Philadelphia, Pa.), Vol. 4, Issue 8, pp. 1209-21, (2011) (PubMed).

Target Details

Target: Glucosidase, beta

Alternative Name: beta-Glucosidase (Glucosidase, beta Products)

Synonyms: BETA-GLUCOSIDASE Kit, GLUC Kit, T12J13.8 Kit, T12J13_8 Kit, beta glucosidase 25 Kit, PSPTO3318 Kit, PSPTO4290 Kit, beta glucosidase 25 Kit, beta-glucosidase Kit, Beta-glucosidase Kit, glucosylceramidase Kit, bglX-2 Kit, BGLU25 Kit, PSPTO_3318 Kit, bglX Kit, bglA4 Kit, LOC100399524 Kit, bglA.2 Kit, bglX-2 Kit

Background: Quantitative determination of beta-glucosidase activity by colorimetric (405nm) method.
Procedure: 20 min.

beta-glucosidase is a glucosidase enzyme which acts upon beta1->4 bonds linking two glucose or glucose-substituted molecules (i.e., the disaccharide cellobiose). beta-glucosidases are required by organisms (some fungi, bacteria, termites) for consumption of cellulose. Lysozyme is also a beta-glucosidase and is present in tears to prevent bacterial infection of the eye. In humans, lower activity of a beta-glucosidase isoform (lysosomal glucocerebrosidase) has been related to Gaucher's disease and Parkinson's disease.
Simple, direct and automation-ready procedures for measuring beta-glucosidase activity are becoming popular in Research and Drug Discovery. This beta-Glucosidase Assay Kit is designed to measure beta-glucosidase activity directly in biological samples without pretreatment. The improved method utilizes p-nitrophenyl-beta-D-glucopyranoside that is hydrolyzed specifically by beta-glucosidase into a yellow colored product (maximal absorbance at 405nm). The rate of the reaction is directly proportional to the enzyme activity.