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Description
Overview
The Glucose Assay kit provides direct measurement of glucose in various biological samples (e.g. serum, plasma, body fluids). Glucose enzyme mix oxidizes glucose to generate a product which reacts with a dye to produce color and fluorescence. The generated color or fluorescence is proportional to the amount of glucose in the sample.
Catalogue Number
CBA086
Brand Family
Calbiochem®
Application Data
References
References
Lee, Y.S. et al. 2007.Diabetes56, 1671.
Product Information
Detection method
Colorimetric or Fluorimetric
Form
100 tests
Format
96-well plate
Kit contains
Glucose Assay Buffer, Glucose Probe, Glucose Enzyme Mix, and Glucose Standard, and a user protocol.
Harmful in contact with skin and if swallowed. Irritating to eyes and skin.
S Phrase
S: 24/25-26-36-45
Avoid contact with skin and eyes. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable protective clothing. In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible).
Product Usage Statements
Intended use
The Glucose Assay Kit provides direct measurement of glucose in various biological samples (e.g., serum, plasma, body fluid, food, growth medium, etc.). The method is rapid, simple, sensitive, and suitable for high throughput. The assay is also suitable for monitoring glucose levels during fermentation and glucose feeding in protein expression processes.
Storage and Shipping Information
Ship Code
Dry Ice Only
Toxicity
Multiple Toxicity Values, refer to MSDS
Storage
-20°C
Storage Conditions
Upon arrival store the entire contents of the kit at (-20°C). Protect from light. Note: Allow reagents warm to room temperature and briefly centrifuge vials before opening.
Do not freeze
Ok to freeze
Mercury prohibited
To comply with ban of sale of mercury-added products required by The Interstate Mercury Education and Reduction Clearinghouse (IMERC), this product is prohibited to be sold in the following US states: Rhode Island and Connecticut.
Packaging Information
Transport Information
Supplemental Information
Kit contains
Glucose Assay Buffer, Glucose Probe, Glucose Enzyme Mix, and Glucose Standard, and a user protocol.
Upon arrival store the entire contents of the kit at (-20°C). Protect from light. Note: Allow reagents warm to room temperature and briefly centrifuge vials before opening.
Intended use
The Glucose Assay Kit provides direct measurement of glucose in various biological samples (e.g., serum, plasma, body fluid, food, growth medium, etc.). The method is rapid, simple, sensitive, and suitable for high throughput. The assay is also suitable for monitoring glucose levels during fermentation and glucose feeding in protein expression processes.
Background
Glucose (C6H12O6; FW: 180.16) is a very important fuel source for generating universal energy molecules ATP. Glucose level is a key diagnostic parameter for many metabolic disorders. Measurement of glucose can be very important in both research and drug discovery processes.
Principles of the assay
The Glucose Assay Kit utilizes the Glucose Enzyme Mix to specifically oxidize glucose to generate a product that reacts with a dye to generate color (&lambda = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The generated color and fluorescence is proportional to the glucose amount.
Prepare test samples in a total volume of 50 µl/well with Glucose Assay Buffer. Serum can be directly diluted in Glucose Assay Buffer; typically 0.5-2 µl/assay is sufficient (normal serum contains ~5 nmol glucose/µl). We suggest testing several doses of your sample to make sure the readings are within the standard curve range.
Reagent preparation
• Glucose Enzyme Mix: Dissolve the Glucose Enzyme Mix in 220 µl Glucose Assay Buffer. Aliquot and store at -20°C. Use within two months. • Glucose Probe: Supplied ready to use. Warm to room temperature to thaw the DMSO. • Standard Curve: For colorimetric assay, dilute the Glucose Standard to 1 nmol/ml by adding 10 µl of the Glucose Standard to 990 µl of Glucose Assay Buffer, mix well. Add 0, 2, 4, 6, 8, 10 µl into each well individually. Adjust volume to 50 µl/well with Glucose Assay Buffer to generate 0, 2, 4, 6, 8, 10 nmol/well of Glucose Standard.
For fluorometric assay, dilute the Glucose Standard solution to 0.1 nmol/µl by adding 10 µl of the Glucose Standard to 990 µl of Glucose Assay Buffer, mix well. Then take 20 µl into 180 µl of Glucose Assay Buffer. Mix well. Add 0, 2, 4, 6, 8, 10 µl into each well individually. Adjust volume to 50 µl/well with Glucose Assay Buffer to generate 0, 0.2, 0.4, 0.6, 0.8, 1.0 nmol/well of the Glucose Standard.
Detailed protocol
1. Prepare reagent mix as indicated below; prepare a volume that is sufficient for the number of assays to be performed; 50 µl per well is required. Each 50 µl of reaction mix should contain the following components: • 46 µl Glucose Assay Buffer • 2 µl Glucose Probe • 2 µl Glucose Enzyme Mix Mix well. 2. Add 50 µl Reaction Mix to each well containing a diluted Glucose Standard or test sample(s). Mix well. 3. Incubate the reaction for 30 min at 37°C, protect from light. 4. Measure the absorbance at 570 nm for a colorimetric assay or at excitation/emission = 535/590 nm for a fluorometric assay using an appropriate microplate reader 5. Correct for background by subtracting the value derived from the 'zero' glucose control from all Standard and sample readings. Note: The background reading can be significant and MUST be subtracted from Standard and sample readings. Glucose concentrations of the test samples can then be calculated based on the standard curve generated.
Standard curve
Figure 1: Standard Curve
Assay Range
1-200 µM
Registered Trademarks
Calbiochem® is a registered trademark of , KGaA, Darmstadt. InteractivePathways™ is a trademark of , KGaA, Darmstadt.
