Description

Invertase (b-fructofuranosidase, EC 3.2.1.26) is an enzyme that catalyzes the hydrolysis of sucrose to fructose and glucose. Invertases cleave at the O-C(fructose) bond, whereas a related enzyme sucrase (EC 3.2.1.48) cleaves[F1] [F2] [F3] [F4] [F5] [F6]  at the O-C(glucose) bond. A wide range of microorganisms produce invertase and can, thus, utilize sucrose as a nutrient. Invertase assay finds wide applications in environmental (e.g. soil), agricultural and food (confectionery) industry.

BioAssay Systems’ Invertase Assay Kit provides a convenient and ultra-sensitive colorimetric and fluorimetric means to measure invertase activity. In the assay, invertase cleaves sucrose, resulting in the formation of fructose and glucose, which is determined by a colorimetric (570nm) or fluorimetric method (l_em/ex = 585/530nm). The assay is simple, sensitive, stable and high-throughput adaptable.

Key features

Safe. Non-radioactive assay.

Sensitive and accurate. As low as 0.007 U/L invertase activity can be quantified[BAS17] .

Homogeneous and convenient. "Mix-incubate-measure" type assay. No wash and reagent transfer steps are involved.

Robust and amenable to HTS: can be readily automated on HTS liquid handling systems for processing thousands of samples per day.

Applications

Invertase and sucrase activity determination in biological and environmental (e.g. soil) samples.

Evaluation and screening for invertase inhibitors.

Kit Contents

Storage conditions: store 10x Reaction Buffer and Assay Buffer at 4°C and other reagents at -20°C. This product is shipped on ice. Shelf life of 3 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.

Assay Procedure

Interference: thiols (b-mercaptoethanol, dithioerythritol etc) at > 10 mM interfere with this assay and should be avoided. Glucose, if present in the sample, should be removed by dialysis or membrane filtration.

1. Assay Preparation. Prior to assay, bring all components to room temperature, briefly centrifuge tubes before opening. Dilute the provided 10x Reaction Buffer and 10x Sucrose to 1-fold by mixing 1 vol of the reagent with 9 vol of dH₂O. Use the diluted reagents for all assays.

     For glucose standard curve, mix 5 mL Glucose Standard with 828 mL dH₂O (final 100 mM). Dilute as follows  and  transfer 40 mL standards