Nutrient metabolism is necessary for energy production in cells and regulates various cellular functions, including gene expression. Glucose is one of the key substrates for the generation of ATP and sustaining cellular homeostasis. Thus, glucose metabolism has been the subject of intense investigations. In cancer research, tumor cells enhance glucose uptake and consumption for their growth and proliferation. Therefore, elevated glucose uptake is a marker of tumors, and glucose transporters are important targets in cancer treatment.
One standard method for evaluating the glucose uptake ability of cells uses radioisotope-labeled glucose. Although this method has been used for many years, it requires special handling facilities and disposal of radioactive materials. The enzyme cycling method using 2-deoxy-D-glucose, which enables colorimetric and fluorometric plate assays, cannot be applied to cell imaging and flow cytometry. Recently, 2-NBDG, a fluorescently labeled glucose analog, has been used widely to detect cellular glucose uptake by fluorescence imaging and flow cytometry1). However, the sensitivity of this method is poor because of the low fluorescence intensity of 2-NBDG.
To resolve these limitations, novel fluorescent probes, Glucose Uptake Probes, were developed. These probes emit strong fluorescence, allowing highly sensitive detection of cellular glucose uptake by fluorescence imaging, flow cytometry or microplate assay. The WI Solution in these kits enhances cellular retention of the probe to give more reliable data.
Glucose Uptake Assay Products
| Products | Plate Reader | Microscope / FCM |
|---|---|---|
| Glucose Uptake Assay Kit-Blue | ✗ | ✓ |
| Glucose Uptake Assay Kit-Green | ✓ | ✓ |
| Glucose Uptake Plate Assay Kit (Green) | ✓ Best* | ✗ |
| Glucose Uptake Assay Kit-Red | ✓ | ✓ |
*No washing step, minimizing cell stress and detachment while improving reproducibility.
Technical info
Four Advantages Compare to Existing Method
Glucose Uptake Probe-Red allowing highly sensitive detection of cellular glucose uptake by fluorescence imaging or flow cytometry. The WI Solution in this kit can enhance cellular retention to provide more reliable experimental data. Also, compare with the existing method (2-NBDG), the measurement time can be significantly reduced.
①Multi-color Imaging
Glucose Uptake Probe can easily co-stain with other fluorescent reagents. The following image is a co-staining result with Glucose Uptake Probe-Red and Lipid Droplets (Green: Lipi-Green, Code LD02) in adipocytes differentiated 3T3-L1 cells.
Cell line: 3T3-L1
Imaging Conditions
Glucose Uptake Probe-Red: Ex=533-557 nm, Em=570-640 nm
Lipi-Green: Ex=450-490 nm, Em=500-550 nm
②Plate Reader Compatibility
Besides fluorescent microscopy and Flow Cytometer, Glucose Uptake Probe-Red is also compatible with Plate Reader. The following data is separately using High Glucose medium and No Glucose medium cultured A549 cells to confirm the uptake inhibition of Glucose Uptake Probe-Red.
Cell Line: A549
Experimental Conditions:
Glucose Uptake Assay Kit-Red: Ex=545 nm, Em=605 nm
③Quick 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.
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④Reduces Dye Leakage After Staining
The WI Solution in this kit can enhance cellular retention to provide more reliable experimental data.
For more details, please refer to the page of (Glucose Uptake Assay Kit-Green, Code: UP02).
Comparison with Existing Method
The comparison of the Glucose Uptake Probe Series and the existing method(2-NBDG) is as below.
| product name | Fluorescence microscope | Plate reader detection | FCM detection | Retention ability | Fluorescence characteristics |
|---|---|---|---|---|---|
| Glucose Uptake Assay Kit-Blue | ✓ | – | ✓ | 1 hour * | λex:386 nm λem:474 nm |
| Glucose Uptake Assay Kit-Green | ✓ | ✓ | ✓ | 1 hour * | λex:507 nm λem:518 nm |
| Glucose Uptake Assay Kit-Red | ✓ | ✓ | ✓ | 1 hour * | λex:560 nm λem:572 nm |
| 2-NBDG | ✓ | – | ✓ | 30 minutes or less * | λex:465 nm λem:540 nm |
*Result of A549 cells, the retention time for other cell lines may be different.
Experimental Example: Inhibition of Glucose Uptake by Cytochalasin B
Using Glucose Uptake Probe Series to detect the inhibition of glucose uptake by the glucose transporter inhibitor Cytochalasin B in HepG2 cells with high sensitivity.
Imaging with microscopy
(Scale bar: 50 μm)
＜Experimental Conditions＞
Cell Line: HepG2
Miedum: MEM （5.5 mmol/l Glucose）
Incubation Conditions: 5 µmol/l Cytochalasin B / MEM （5.5 mmol/l Glucose, 10% FBS）, 37℃, 24 hr
Reagent Conditions: Glucose Uptake Probe (500x dilution)/DMEM （0 mol/l Glucose）, 37℃, 15 min
Device: Fluorescent Microscopy
Glucose Uptake Assay Kit-Blue: Ex=340-380 nm, Em: 435-485 nm
Glucose Uptake Assay Kit-Green: Ex=450-490 nm, Em: 500-550 nm
Glucose Uptake Assay Kit-Red: Ex=533-557 nm, Em: 570-640 nm
Experimental Example: Enhanced Glucose Uptake by Insulin
Using Glucose Uptake Probe to evaluate insulin-induced glucose uptake enhancing in adipocytes with high sensitivity.
Imaging with microscopy
(Scale bar: 50 µm)
＜Experimental Conditions＞
Cell: mouse adipocyte
Miedum: DMEM (5.5 mmol/l Glucose, 10% FBS)
Inducer Conditions：0 or 1 µmol/l Insulin / DMEM (0 mmol/l Glucose , serum-free), 37℃, 15 min
Reagent Conditions：Glucose Uptake Probe (500x dilution) /DMEM (0 mmol/l Glucose, serum-free), 37℃, 15 min
Device: Fluorescent Microscopy
Glucose Uptake Assay Kit-Blue: Ex=340-380 nm, Em: 435-485 nm
Glucose Uptake Assay Kit-Green: Ex=450-490 nm, Em: 500-550 nm
Glucose Uptake Assay Kit-Red: Ex=533-557 nm, Em: 570-640 nm
Difference from Glucose Assay Kit
The difference between Glucose Uptake Assay Kit and Glucose Assay Kit-WST(Code: G264)
1. Glucose Assay Kit-WST can quantify the glucose concentration in cell supernatants.
Glucose Uptake Assay Kit does not apply for the quantification of glucose.
2. Glucose Uptake Assay Kit can evaluate cellular glucose uptake capability in a short time.
Glucose Assay Kit-WST can not measure the glucose level changes in a short period of time.
For more details, please refer to the page of Glucose Uptake Assay Kit-Green (Code: UP02).
