Glucose Uptake Assay Kits

Glucose Uptake Assays

Measure physiological glucose uptake

Assay Genie's Glucose Uptake Assays are simple, ultra-sensitive and easy to use kits that allow researchers to study the mechanism of glucose uptake by cells. The glucose uptake assay provides a direct, powerful tool for studying this process, as well as for screening and characterization of drugs that regulate glucose uptake during normal health and disease development.

What is glucose and why is studying glucose uptake important?

Glucose is a ubiquitous energy source in most organisms and plays a pivotal role in cellular metabolism and homeostasis. Glucose uptake is one of the key processes for cellular glucose metabolism. The study of glucose uptake can provide important information for understanding glucose metabolism and regulation in normal and disease development such as diabetes.

Figure 1: Glucose uptake in Jurkat and HeLa cells. 2.5x105 Jurkat cells were treated with or without 4 μl phloretin (1X concentration) for 45 min. After treatment, cells were washed and incubated with Glucose Uptake Reagent, glucose uptake enhancer and the same concentration of phloretin for another 30 minutes according to the kit protocol. A) Comparison of histogram from the flow analysis showing the inhibition of glucose uptake by phloretin in Jurkat cells (Black: negative control cells; orange: in the presence of phloretin; blue: without phloretin). B) Images of Jurkat cells obtained using fluorescent microscope (Top: treated with phloretin; Bottom: without phloretin treatment). C) Glucose Uptake in HeLa cells: HeLa cells showing uptake of glucose uptake reagent in the cytoplasm. Cells were strained with glucose uptake reagent for 30 minutes and fixed. Image was taken using a fluorescent microscope with a 60X objective lens.

Glucose uptake

Glucose uptake has a variety of methods and transporters, and depends upon the metabolic demand of the cell type and availability of glucose.

There are over ten different facilitated diffusion glucose transporters which transports glucose down its concentration gradient without ATP hydrolysis. In the kidneys, secondary active transport is used to uptake Glucose against its concentration gradient to ensure that very little glucose is excreted in urine.

Assay Genie Direct Glucose Uptake Assay Principle

Assay Genie's Direct Glucose Uptake Assay kit is highly sensitive, safe and easy to use. A specific hexokinase inhibitor that inhibits hexokinase, the first enzyme metabolizing glucose in cells is used to arrest glucose consumption after its uptake.

Glucose Uptake is measured by using a set of enzymatic reactions that specifically oxidize glucose producing intermediates that react with the OxiGenie Probe generating a fluorescence signal (Ex/Em=535/587 nm). The fluorescence signal is directly proportional to the amount of glucose that has been taken up and accumulated inside the cells.

Features & Benefits

  • Real direct results - Rapidly quantify direct glucose uptake using a specific glucose detection enzyme.
  • Performance - Less than 15% assay variability versus 60% with 2-DG based methods.
  • Sensitive - Measure down to 5pmol per well.
  • Easy-to-use - Simple protocol with 6 components.
  • Safe - Non-radioactive method of detecting glucose uptake in cells.

Figure 2. A) Glucose standard curve. B) Glucose Uptake time course, Jurkat Cells: Cells were starved (Glucose-free, FBS-free media, inhibitor incubation time: 2 h. C) 3T3-L1 cells were Glucose and FBS Deprived for 24 hrs, switched to media with Glucose, stimulated without or with Insulin (10 ng/ml) for 15 min (green bars), with or without 1X Inhibitor (pink bars). D) HeLa cells were Glucose and FBS deprived for 2 hr, then switched to Glucose and FBS-free media (Control), or complete media (10% FBS) with or without 1X Inhibitor for 30 min (Blue bars: no inhibitor; teal bars: with inhibitor).

Other tools for measuring glucose uptake

- 2-deoxyglucose (2-DG) - 2-DG has been widely used in glucose uptake assays because of its structural similarity to glucose. As with glucose, 2-DG can be taken up by glucose transporters and metabolized to 2-DG-6-phosphate (2-DG6P). 2-DG6P, however, cannot be further metabolized, and thus accumulates in the cells. The accumulated 2-DG6P is directly proportional to 2-DG (or glucose) uptake by cells. To validate the assay, the kit includes phloretin, a natural phenol that inhibits glucose uptake. This easy-to-use, non-radioactive kit allows imaging and accurate measurement of glucose uptake in cultured cells in response to insulin, growth factors etc. Such kits can either be colorimetric or fluorometric in nature.

    • Glucose Uptake Colorimetric Assay (BN00900) - The 2-DG6P is oxidized to generate NADPH, which can be determined by an enzymatic recycling amplification reaction. This kit can detect glucose uptake as low as 10 pmol/well.

Figure 3. (A) 2-DG6P Standard curve (B) 2-DG uptake in 3T3-L1, human adipocyte and Hela cells. To scale on the same graph, data from 3T3-L1 cells is plotted at 10% of true value. Assays were performed following kit protocol. 2-DG = 2-deoxyglucose, I = Insulin; P = Phloretin.

    • Glucose Uptake Fluorometric Assay (BN00890) - The accumulated 2-DG6P is enzymatically oxidized and coupled to Assay Genie's PicoProbe, which generates fluorescence in the presence of NADPH. This kit can detect glucose uptake as low as 50 pmol/well and can be used for a variety of cell types.

Figure 4. (A) 2-DG6P Standard curve (B), (C)  and (D) 2-DG uptake in human adipocytes, Hela Cells and 3T3-L1 cells respectively. Assays were performed following kit protocol. I=Insulin; P=Phloretin.

- 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]-D-glucose (2-NBDG) (BN00906)- 2-NBDG is a fluorescent deoxyglucose analog that can be taken up by cells through glucose transporters. However, 2-NBDG cannot be fully utilized in glycolysis because of its modification and thus accumulates inside the cells. Fluorescence generated by this fluorescent glucose analog is proportional to glucose uptake by the cells and can be used to measure glucose uptake using fluorescent microscopy and flow cytometry.

Figure 5. Glucose uptake in Jurkat and HeLa cells. 2.5x105 Jurkat cells were pretreated with or without 4 µl phloretin (1X concentration) for 45 min. After pretreatment, cells were washed and incubated with 2-NBDG Reagent, Glucose Uptake Enhancer, and the same concentration of phloretin for another 30 min. according to kit’s protocol. (A) Comparison of histograms from flow analysis showing the inhibition of glucose uptake by phloretin in Jurkat cells (Black: negative control cells; orange: in the presence of phloretin; blue: without phloretin).

Tools for glucose uptake measurement

Assay Code Read-out Sensitivity Sample Type Pack Size

BN01098

Fluorometric

N/A

Adherent/Suspension

50 Assays

BN00908

Fluorometric

N/A

Adherent/Suspension

50 Assays

BN00906

Fluorometric

N/A

Adherent/Suspension

50 Assays

BN00905

Fluorometric

N/A

Adherent/Suspension

50 Assays

BN00900

Colorimetric

10 pmol

Adherent/Suspension

100 Assays

BN00890

Fluorometric

100 pmol

Adherent/Suspension

100 Assays