Glucose Uptake Assay Kit (DTNB)
Glucose metabolism is a process that converts ingested glucose into energy, which is the main source of energy supply in living organisms. Studies have shown that glucose metabolism is closely related to many diseases such as cancer, diabetes and obesity, so it is important to monitor glucose uptake.
This kit is designed for the detection of glucose uptake based on the DTNB chromogenic reaction. The glucose analog 2-deoxyglucose (2-DG) is taken up into cells via glucose transporters and is subsequently phosphorylated to 2-deoxyglucose-6-phosphate (2-DG6P) by endogenous kinases. Glucose-6-phosphate dehydrogenase (G6PDH) then catalyzes the conversion of 2-DG6P, while simultaneously reducing NADP+ to NADPH. In the presence of glutathione reductase (GR), NADPH reduces oxidized glutathione (GSSG) to reduced glutathione (GSH). The generated GSH then reacts with the chromogenic substrate DTNB to produce yellow-colored TNB, which has a maximum absorbance at 412 nm. The absorbance at 412 nm is directly proportional to the cellular glucose uptake capacity.
This kit enables sensitive, rapid, and non‑radioactive measurement of glucose uptake. It has been validated to exhibit a good linear relationship within the range of 1 to 100 pmol. Compared with the WST‑8 Glucose Uptake Assay Kit (Cat. No. K2303), this kit offers higher sensitivity and is therefore more suitable for samples with relatively low glucose uptake activity.
Figure 1: Typical standard curve and assay data. (A) The reference standard curve, (B) the glucose uptake capacity of HeLa cells after oleic acid induction. Insulin(I), which is used to enhance glucose uptake; Phloretin(P), which is used to inhibit glucose uptake.
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Components |
100 Assays |
500 Assays |
Storage |
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Glucose Uptake Lysis Buffer |
15 mL |
70 mL |
-20°C |
|
Glucose Uptake Assay Buffer |
10 mL |
50 mL |
-20°C |
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KRPH Buffer |
12 mL |
60 mL |
-20°C |
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Enzyme Solution A |
500 μL |
2.5 mL |
-20°C |
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Reaction Mix |
200 μL |
1 mL |
-20°C |
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Stop Buffer |
600 μL |
3 mL |
-20°C |
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GR Buffer |
600 μL |
3 mL |
-20°C |
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Recycling Mix |
200 μL |
1 mL |
-20°C |
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Enzyme Solution B (15X) |
20 μL |
100 uL |
4°C |
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DTNB (10X) |
220 μL |
1.1 mL |
-20°C away from light |
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2-DG (10 mM) |
1 mL |
5 mL |
-20°C |
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2-DG6P Standard (1 mM) |
20 μL |
60 μL |
-20°C |
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Shipping: Blue ice Shelf life: 1 year |
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