Glucose Uptake-Glo™ Assay
A Simple, Non-Radioactive Assay to Measure Glucose Uptake Inside Cells
- Homogeneous protocol
- Achieve sensitivity with broad linearity
- Reliable and reproducible results
Catalog Number:
Size
Catalog Number: J1341
Catalog Number: J1342
Catalog Number: J1343
Measuring Glucose Uptake
Glucose metabolism is a key process in many organisms. A lack of insulin-stimulated glucose uptake is associated with type 2 diabetes, while high glucose uptake is a sign of the high glycolytic rates associated with cancer. Measuring glucose uptake can determine the effects of various treatments for diabetes and cancer.
The gold standard method of assaying glucose uptake relies on detection of radio-labeled glucose analogs. Alternative colorimetric and fluorometric glucose uptake assays often lack the sensitivity and robustness needed to reliably measure glucose uptake in cells.
シンプルでNon-RI のグルコース取込みアッセイ
Glucose Uptake-Glo™ Assay は細胞でのグルコース取込みを発光法により測定するプレートベースのホモジニアスアッセイで、2-deoxyglucose-6-phosphate (2DG6P) の検出がベースになっています。
"Promega products are reliable and can be purchased without any hesitation. My experiment went very well as expected and we have more plans to conduct uptake experiments using the Glucose Uptake-Glo™ Assay."
Glucose Uptake-Glo と従来のRI 法との比較
2-デオキシグルコース (2DG) を細胞に添加するとグルコースと同様に細胞膜を透過し、迅速にリン酸化され、2-deoxyglucose-6-phosphate (2DG6P) を生成します。Stop Buffer を加えると2DGの輸送が停止し、細胞を溶解し、細胞内に残存するNADPHを破壊してタンパク質を不活性化します。Neutralization Buffer を添加すると 2DG6P Detection Reagent 添加前の溶液が中和されます。試薬に含まれる glucose-6-phosphate dehydrogenase (G6PDH) は 2DG6P を酸化して6-phosphodeoxygluconate (6PDG) を生じ、NADP+を還元してNADPHにします。レダクターゼはNADPHを用いてルシフェリン前駆体から発光酵素ルシフェラーゼの基質ルシフェリンに変換します。
がんおよび脂肪細胞株における代謝変化の測定
MCF7 がん細胞
がんモデルでは細胞が酸素不足になると、この低酸素条件により細胞内代謝が酸化的リン酸化から解糖系にシフトします。この結果としてグルコースの取込みが増加します。
MCF7 cells grown under hypoxia (1% oxygen) show an increase in Glucose Uptake-Glo™, indicating an increased glycolytic rate. The same cells demonstrate no significant change in viability using the RealTime-Glo™ and CellTiter-Fluor™ Assays.
脂肪細胞
脂肪細胞モデルを用いて、3T3L1-MBX 脂肪細胞のグルコース取込み量の変化が細胞健全性に対して顕著な影響を与えなかったことを示しました。Glucose Uptake-Glo™ Assay と RealTime-Glo™ Assay のマルチアッセイにより細胞健全性に対する影響とグルコース取込み変化をすぐに分離して測定することができた。
Results of the Glucose Uptake-Glo™ Assay when adipocyte cells are treated under various conditions, where "None" indicates the basal level of glucose uptake. Insulin induces translocation of glucose transporters to the cell surface, and thus increases glucose uptake above basal levels. Cytochalasin B is a glucose transporter inhibitor which decreases glucose uptake. LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, is an essential insulin signaling enzyme which decreases glucose uptake relative to insulin alone.
Results of the RealTime-Glo™ Assay when adipocyte cells are treated under various conditions, where "None" indicates the basal level of glucose uptake. Although there are dramatic changes in glucose uptake, as seen in the accompanying panel, there are minimal changes in viability as measured by the RealTime-Glo™ Assay.
FAQs
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Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Luciferase Reagent |
J151A | 1 × 5ml |
Stop Buffer |
J152A | 1 × 15ml |
Neutralization Buffer |
J153A | 1 × 15ml |
NADP+ |
J154A | 1 × 50μl |
G6PDH |
J155A | 1 × 125μl |
2DG |
J156A | 1 × 250μl |
2DG6P Standard |
J157A | 1 × 50μl |
Reductase |
G884C | 1 × 25μl |
Reductase Substrate |
G885A | 1 × 55μl |
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
Luciferase Reagent |
J151B | 1 × 10ml | |
Stop Buffer |
J152A | 1 × 15ml | |
Neutralization Buffer |
J153A | 1 × 15ml | |
NADP+ |
J154B | 1 × 100μl | |
G6PDH |
J155B | 1 × 250μl | |
2DG |
J156A | 1 × 250μl | |
2DG6P Standard |
J157A | 1 × 50μl | |
Reductase |
G884A | 1 × 55μl | 6mg/ml |
Reductase Substrate |
G885A | 1 × 55μl |
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
Luciferase Reagent |
J151C | 1 × 50ml | |
Stop Buffer |
J152A | 1 × 15ml | |
Neutralization Buffer |
J153A | 1 × 15ml | |
NADP+ |
J154C | 1 × 500μl | |
G6PDH |
J155C | 1 × 1.25ml | |
2DG |
J156A | 1 × 250μl | |
2DG6P Standard |
J157A | 1 × 50μl | |
Reductase |
G884B | 1 × 275μl | 6mg/ml |
Reductase Substrate |
G885A | 1 × 55μl |
U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.
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