alpha-Glucosidase Assay Kit

N° du produit ABIN1000246

Aperçu rapide pour alpha-Glucosidase Assay Kit (ABIN1000246)

Antigène: AGLU (alpha-Glucosidase (AGLU))

Application: Activity Assay (AcA)

Détail du produit

Gamme de detection: 2-250 U/L

Seuil minimal de détection: 2 U/L

Attributs du produit: 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.

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

Matériel non inclus: Pipeting devices and accessories (e.g. multi-channel pipettor). Clear bottom 96-well plates (e.g. Corning Costar) and plate reader.

Information d'application

Indications d'application: Direct Assays: beta-glucosidase activity in biological samples.
Characterization and Quality Control for beta-glucosidase production.
Drug Discovery: high-throughput screen and evaluation of beta-glucosidase inhibitors.

Protocole: 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.

Préparation des réactifs:

Equilibrate reagents to room temperature. The Working Reagent 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.

Préparation de l'échantillon:

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, Triton X-100, Tween, digitonin, EDTA and Tris.

Restrictions: For Research Use only

Stockage

Stock: 4 °C

Détail du antigène

Antigène: AGLU (alpha-Glucosidase (AGLU))

Autre désignation: alpha-Glucosidase

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

alpha-glucosidase hydrolyzes the terminal, non-reducing 1,4-linked alpha-D- glucose residues with release of alpha-D-glucose. alpha-glucosidase is needed by all animals to hydrolyze maltose to glucose for use as a food. Aberrant activities have been implicated in diseases such as diabetes and Pompe disease.
Simple, direct and automation-ready procedures for measuring alpha-glucosidase activity are becoming popular in Research and Drug Discovery. The alpha-Glucosidase Assay Kit is designed to measure alpha-glucosidase activity directly in biological samples without pretreatment. The improved method utilizes p-nitrophenyl-alpha-D-glucopyranoside that is hydrolyzed specifically by alpha-glucosidase into a yellow colored product (maximal absorbance at 405nm). The rate of the reaction is directly proportional to the enzyme activity.