CheKine Glucose Oxidase Activity (GOD) Assay Kit | KTB1310

 The glucose oxidase enzyme (GOx) (EC 1.1.3.four) is a dimeric enzyme that catalyzes oxidation of beta-D-glucose into hydrogen peroxide and D-glucono-1,five-lactone, that’s hydrolyzed to gluconic acid. This enzyme is produced through sure species of insects, fungi, and micro organism. Glucose oxidase is broadly used for the willpower of glucose in frame fluids and in casting off residual glucose and oxygen from beverages, meals and different agricultural products. Furthermore, Glucose oxidase is normally used in biosensors to come across glucose.

CheKine Glucose Oxidase Activity (GOD) Colorimetric Assay Kit presents a easy and easy colorimetric assay for the take a look at of Glucose Oxidase hobby in loads of organic samples along with serum, plasma, animal tissues, cells, micro organism, organic fluids along with Urine. The precept is that GOD hobby is decided through a coupled enzyme assay, wherein GOD oxidizes D-glucose ensuing withinside the manufacturing of hydrogen peroxide (H2O2) that reacts with chromogen in an acid condition, produce a product that may be measured at OD580 nm. Therefore, the glucose oxidase hobby gift withinside the pattern is proportional to the sign obtained.

Materials Required however Not Supplied

• Microplate Reader able to measuring absorbance at OD580 nm ·
• Incubator, Ice Maker, Refrigerated Centrifuge ·96-properly Plate with clean flat bottom, precision Pipettes, Disposable Pipette Tips ·Deionized Water ·
• Dounce homogenizer (for Tissue Samples)

Reagent Preparation

Assay Buffer (1×): Equilibrate to room temperature earlier than use.Make a 1:20 dilution of the focused Assay Buffer (20×) with deionized water. This very last Assay Buffer must be used to dilute the H2O2 requirements and samples previous to assay.When saved at four°C, this diluted Assay Buffer (1×) is solid for at the least months.

Glucose (0.2 M): Ready to apply as supplied. Equilibrate to room temperature earlier than use. Please aliquot the unused reagents and save at -20℃. H2O2

Standard (0.88 M): Ready to apply as supplied. Equilibrate to room temperature earlier than use, included from mild throughout the assay. Please aliquot the unused reagents and save at -20℃. Chromogen: Ready to apply as supplied. Equilibrate to room temperature earlier than use, included from mild throughout the assay. Please aliquot the unused reagents and save at -20℃.

Glucose Oxidase (control): The vial incorporates Glucose Oxidase from Aspergillus niger, that’s used as a superb control. Take 2 μL of the Glucose Oxidase and dilute with 998 µL of Assay Buffer, included from mild on ice throughout the assay. Please aliquot the unused reagents and save at -20℃.

Standard preparation: Prepare 10 mM of H2O2 Standard through diluting 10 µL 0.88 M H2O2 Standard into 870 µL 1×Assay Buffer. Prepare forty µM of H2O2 Standard through diluting four µL 10mM H2O2 Standard into 996 µL1×Assay Buffer. Using forty µM trendy, put together trendy curve dilution.

Sample Preparation

1. Animal Tissue samples: Perfuse tissue with bloodless PBS to dispose of any crimson blood cells. Homogenize tissue at 1 mL/0.1 g in bloodless 1 ×Assay Buffer. Centrifuge at 12,000 g for five min at four℃. Use supernatant for assay, hold on ice throughout the assay.

2. Cell and micro organism: Harvest the quantity of cells or micro organism essential for every assay (preliminary recommendation=five×106 cells/assay). Wash cells with bloodless PBS. Homogenize cells (five×106 ) with 1 mL of ice-bloodless 1×Assay Buffer. Centrifuge at 12,000 g for five min at four℃. Use supernatant for assay, hold on ice throughout the assay.

3. Plasma, Serum, Urine (and different organic fluids): Tested immediately through including samples to microcentrifuge tubes. However, to find the premier values and make sure your readings will fall withinside the trendy values, we advocate acting numerous dilutions of the samples.

Note: Fresh samples are advocated. If now no longer assayed immediately, samples may be saved at -80℃ for one month. If the protein attention of the pattern is want to decided, it’s miles advocated to apply Abbkine Cat #: KTD3001 Protein Quantification Kit (BCA Assay) to degree the protein attention of the pattern.