Contact distributor nearby
Need our help ?


Glucose Uptake Assay Kit-Blue

Item # Unit Size
UP01-10
1 set

For Research Use Only Products

Contents: 1 set
・Glucose Uptake Probe-Blue ×1
・WI Solution (50×)  5 ml ×1

The general number of usable assays per 1 set
・35 mm dish x 12
・96-well plate x 1

Storage Condition: Store at 0-5oC
Shipping Condition: Ambient Temperature

MSDS

Detection Principle

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 to sustain 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 common 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 cytometry. However, the sensitivity of this method is poor because of the low fluorescence intensity of 2-NBDG.

To resolve these limitations, a novel fluorescent probe, Glucose Uptake Probe-Blue, was developed. This probe emits strong blue fluorescence (λex = 386 nm, λem = 474 nm), allowing highly sensitive detection of cellular glucose uptake by fluorescence imaging, flow cytometry. The WI Solution in this kit enhances cellular retention of the probe to give more reliable data.



 
Fluorescence Properties

Glucose Uptake Probe-Blue




Please refer to the link below for details regarding several colors of glucose assay kits.


Many researchers are using Dojindo’s Glucose Uptake Assay Kit to evaluate glucose uptake capacity.
Four Advantages Compare to Existing Method

Glucose Uptake Probe-Blue 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 Green or red or any other long-wavelength fluorescent reagent. The following image is a co-staining result with Glucose Uptake Probe-Blue and Lipid Droplets (Red: Lipi-Red, Code LD03) in adipocytes differentiated 3T3-L1 cells.



<Experimental  Conditions>
Cell line: 3T3-L1
Imaging Conditions
Glucose Uptake Probe-Blue: Ex=340-380 nm, Em=435-485 nm
Lipi-Red: Ex=533-557 nm, Em=570-640 nm


②Microscopy & FCM Compatibility

Glucose Uptake Probe-Blue is compatible with microscope and Flow Cytometer. The result below is using Glucose  Uptake Probe-Blue to evaluate A549 cells' glucose uptake capability with No Glucose and High Glucose medium.



Cell Line: A549
Imaging Conditions:
Glucose Uptake Assay Kit-Blue: Ex=340-380 nm, Em=435-485 nm

 


Cell Line: A549
Experimental Conditions:
Glucose Uptake Assay Kit-Blue: Ex=405 nm, Em=425-475 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.

 



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).

 
Green fluorescently labeled glucose analogs, such as 2-NBDG, have been widely used to detect cellular glucose uptake. However, the fluorescence intensity is low, and it is poorly suited for experiments with samples that emit green autofluorescence.
As an alternative, a coumarin-based glucose analog emitting blue fluorescence is also commercially available. However, the sensitivity is poor because of the low fluorescence intensity.
Dojindo’s Glucose Uptake Probe-Blue enables highly sensitive measurement of glucose uptake due to its strong blue fluorescence emission.

 
Comparison Between Glucose Uptake Probe-Blue and Reagent C Emitting Blue Fluorescence

[Protocol]
1. A549 cells/MEM/ 96 well microplate (15,000 cells/well), 37℃, 1day
2. DMEM (0 mM glucose) wash x 2
3. DMEM (0 mM glucose), 37℃, 15 min
4. Glucose Uptake Probe-Blue or Reagent C/DMEM (0 or 25 mM Glucose), 37℃, 15 min
5. WI solution (4℃) wash x 2,
6. WI solution (4℃), r.t., 5 min
7. Remove the supernatant, add WI solution (4℃)
8. Imaging



Imaging of glucose uptake in A549 was performed with either Glucose Uptake Probe-Blue or Reagent C. The cells stained with Glucose Uptake Probe-Blue showed high fluorescence in the absence of glucose but low fluorescence in the presence of glucose, supporting that the probe can be taken up by the cells via glucose transporters. However, Reagent C gave little fluorescence regardless of the presence or absence of glucose.
Does Glucose Uptake Probe-Blue have selectivity for the type of glucose transporter?

No. Glucose Uptake Probe-Blue is thought to be taken up by cells via multiple glucose transporters. However, there are no detailed data on the selectivity and affinity of this probe for glucose transporters.


Is Glucose Uptake Probe-Blue degraded or metabolized after uptaking by cells?

The structure of Glucose Uptake Probe-Blue is stable and will not be degraded during any steps of the protocol. The glucose moiety of Glucose Uptake Probe-Blue may undergo phosphorylation by hexokinase due to its structure, but no further metabolism is expected to occur.

After Glucose Uptake Probe-Blue is uptaken by cells, can the cells be fixed?"

No. Glucose Uptake Probe-Blue leaks from cells after fixation.

Is that possible to store the working solution for a long time?"

No, the Probe working solution cannot be stored. The Probe Stock solution can be stored at -20 oC for 1 month.

What should I do if the sensitivity of measurement using Glucose Uptake Probe is low?"

Please consider the probe concentration (×250 to ×1,000) and staining time (15 minutes to 1 hour) as initial considerations.

What should I do if the fluorescence background is high in the measurement using Glucose Uptake Probe-Blue?"

There is a possibility that some of the probes were not taken up by the cells remained. Please wash the cells again with WI solution.

How long is the probe retained in the cells after washing with WI Solution?"

The Probe is retained in the cells for about 1 hour at room temperature in WI Solution. Please note that the retention time may vary depending on the cell type.

Can the Glucose Uptake Assay Kit-Blue be used to quantify glucose in the culture medium or in cells?"

No, please use our product Glucose Assay Kit-WST (product code G264).

Is it possible to quantify Glucose Uptake Probe taken up into cells?"

No, this kit measures the fluorescence intensity of the probe taken up into the cell as an indication of the glucose uptake capacity of the cell.