Glucose Uptake Assay Kit-Green

The use of a high-fluorescence intensity dye enables higher sensitivity measurements in a shorter time than the conventional method (2-NBDG).

1. High-sensitivity determination

Uses a fluorescent dye with superior brightness, which is different from the fluorescence intensity of 2-NBDG, which decreases in water.

Deteciton conditions
Cells: A549 cells
Detection system: Fluorescence microscope
Filter set: GFP (Ex: 470/40 nm, Em: 525/50 nm)

2. Quick process
Significantly reduced measurement time despite using the same process as 2-NBDG.

The operation is very simple, requiring only three steps: pretreatment, staining (incorporation), and washing.

3. Plate reader compatibility
Supports plate reader measurements that are difficult to perform with 2-NBDG.

Deteciton conditions
Cells: A549 cells
Ex: 488 nm, Em: 520 nm

4. Reduces dye leakage after staining
The included WI Solution suppresses reagent leakage and ensures stable data acquisition.

When cells are washed with HBSS

When cells are washed with WI Solution

Deteciton conditions
Cells: A549 cells
Detection system: Fluorescence microscope
Filter set: GFP (Ex: 470/40 nm, Em: 525/50 nm)

Comparison with existing method
Glucose Uptake Probe-Green is compatible with fluorescence microscopes and flow cytometers as well as 2-NBDG. The excitation wavelength is more suitable for 488 nm excitation lasers and filter sets compared with 2-NBDG.

* This is the result of a study using A549 cells, and the leakage time varies depending on the cell type

Difference from Glucose Assay Kit

Difference between Glucose Uptake Assay Kit-Green and Glucose Assay Kit-WST (product code: G264)

1. Glucose Uptake Assay Kit-WST can quantify glucose consumption in cell supernatant. On the other hand, Glucose Uptake Assay Kit-Green cannot quantify glucose.

2. Glucose Uptake Assay Kit-Green can measure the difference of glucose uptake capacity in a short time, while Glucose Assay Kit-WST cannot measure the change of glucose level in a short time.

The difference between the Glucose Assay Kit-WST and this kit will be explained using the following experimental example.

The difference between the Glucose Assay Kit-WST and this kit will be explained using the following experimental example.
Example of experiment: Measurement of glucose consumption and uptake capacity of HepG2 treated with glucose uptake inhibitor (Cytochalasin B).
Flowchart of the experiment and experimental results

Experimental example: Inhibition of glucose uptake by Cytochalasin B
Using this kit, we were able to detect the inhibition of glucose uptake by the glucose transporter inhibitor Cytochalasin B in HepG2 cells with high sensitivity.

Fluorescence microscope observation

(scale bar: 50 μm)

Measurement conditions
Cells: HepG2
Medium: MEM (5.5 mmol/l Glucose)
Culture conditions: 5 µmol/l Cytochalasin B/MEM (5.5 mmol/l Glucose, 10% FBS), 37 °C, 24 h
Staining conditions: Glucose Uptake Probe/DMEM (0 mol/l Glucose) diluted by 500-fold, 37 °C, 15 min
Detection system: Fluorescence microscope
Filter set: GFP (Ex: 470/40 nm, Em: 525/50 nm)

Plate reader detection

Ex: 488 nm, Em: 520 nm

Experimental example: Enhanced glucose uptake by insulin
Using this kit, we were able to measure enhanced insulin-induced glucose uptake in adipocytes with high sensitivity.

(scale bar: 50 µm)

Measurement conditions
Cells: mouse adipocytes
Medium: DMEM (5.5 mmol/l Glucose)
Culture conditions: 1 µmol/l Insulin/DMEM (5.5 mmol/l Glucose), 37 °C, 15 min
Staining conditions: ×500 Glucose Uptake Probe/DMEM (0 mol/l Glucose), 37 °C, 15 min
Detection system: Fluorescence microscope
Filter set: GFP (Ex: 470/40 nm, Em: 525/50 nm)
Scale bar: 50 μm

Plate reader detection

Detection conditions
Ex: 488 nm, Em: 520 nm

* It is difficult to make adipocyte evenly on the wells, so there may be some error in the data.

Experimental procedure
1. Adipocytes were seeded onto ibi 96-well plates and cultured overnight.
2. The cells were washed twice with glucose-free medium DMEM, and then glucose-free medium was added.
3. The cells were incubated at 37 °C for 15 minutes.
4. Probe solution (500-fold dilution) in glucose-free medium was added, and the cells were incubated at 37 °C for 15 minutes.
5. Wash the cells three times with WI Solution (1x) chilled to 4 °C and add WI Solution (4 °C).
6. The cells were observed using a fluorescence microscope and plate reader.

Experimental example: Comparing the glucose uptake capacity of progenitor adipocytes and adipocytes
Using this kit, we were able to detect and quantify the difference in glucose uptake capacity between progenitor adipocytes and adipocytes with high sensitivity.

(scale bar: 50 µm)

Measurement conditions
Cells: preadipocyte, adipocyte
Medium: DMEM (5.5 mmol/l Glucose, 10% FBS)
Staining conditions: Glucose Uptake Probe/DMEM (0 mol/l Glucose) diluted by 500-fold, 37 °C, 15 min
Detection system: Fluorescence microscope
Filter set: GFP (Ex: 470/40 nm, Em: 525/50 nm)

Plate reader detection

Detection conditions
Ex: 488 nm, Em: 520 nm

* It is difficult to make adipocyte evenly on the wells, so there may be some error in the data.

Experimental procedure
1. Progenitor adipocytes and adipocytes were seeded onto ibi 96-well plates and cultured overnight.
2. The cells were washed twice with glucose-free medium DMEM, and then glucose-free medium was added.
3. The cells were incubated at 37 °C for 15 minutes.
4. Probe solution (500-fold dilution) in glucose-free medium was added, and the cells were incubated at 37 °C for 15 minutes.
5. Wash the cells three times with WI Solution (1x) chilled to 4 °C and add WI Solution (4 °C).
6. The cells were observed using a fluorescence microscope and plate reader.