Glucose Assay in Serum

Materials

• FujiFilm Autokit Glucose (Cat 997-03001). Dissolve the Color Reagent in Buffer Solution (150 mL) to make a bottle of 150 mL Color Solution. The solution is stable for 2 weeks at 4C.
• Glucose Standard Solution (200 mg/dL)
• Serum samples (thaw on ice if from -80C)

Procedure

• Plan out 96-well plate, including samples and space for 5 standards and a blank, in duplicates.
• Add 100 uL of Color Solution to each well.
• Add 0, 1, 2, 3, 4, 5 uL to each of the standard wells.
• Add 3 uL of sample to each well.
• Incubate at room temperature for 30 mins (or 5 mins at 37C).
• Read plate at 505 nM and 600 nM.

Calculations

• Use the standard curve to calculate glucose levels.

Reading Plate Instructions

• Turn on the plate reader (switch at the back) and the computer.
• Open ‘Soft Max Pro’.
• Click ‘Settings’.
• Set ‘Wavelengths’ to “2”.
• Set ‘Lm1’ to “490”, ‘Lm2’ to “650”.
• Put plate in, REMOVE THE LID.
• Click ‘Read’.
• Reading: top numbers are 490 uL, bottle numbers are 650 uL.
• To export files: click ‘file’ - ‘export’ - ‘save to computer’ - ‘Bridges’ folder - place in USB.