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| Product Name | Glucose Uptake Cell-Based Assay Kit (2-NBDG) |
| Catalog No. | CMTR-HMM-0053 |
| Description | A fluorescence-based assay that quantifies glucose uptake in live cells using the fluorescent D-glucose analog 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG). 2-NBDG is transported into cells via glucose transporters and is phosphorylated but not further metabolized, leading to intracellular accumulation proportional to glucose uptake rate. |
| Intended Use | Measurement of cellular glucose uptake activity for insulin signaling studies, glucose transporter (GLUT) function research, metabolic phenotyping of cancer and immune cells, and screening of anti-diabetic drug candidates. |
| Principle / Technology | 2-NBDG enters cells through facilitative glucose transporters (GLUT1-GLUT4) and is phosphorylated by hexokinase to 2-NBDG-6-phosphate; unlike native glucose-6-phosphate, 2-NBDG-6-phosphate cannot proceed through glycolysis or the pentose phosphate pathway, resulting in intracellular fluorescent accumulation quantified at Ex/Em 465/540 nm; the signal is proportional to cellular glucose uptake activity over the incubation period. |
| Detection Method | Fluorescence microplate reader (Ex/Em 465/540 nm), fluorescence microscopy with FITC filter set, or flow cytometry (488 nm excitation, FL1 channel) |
| Sample Type | Adherent and suspension mammalian cells, primary adipocytes, skeletal myotubes, insulin-responsive cell lines (3T3-L1 adipocytes, L6 myotubes), cancer cell lines |
| Performance Range / Specifications | 2-NBDG working concentration: 50–150 μM; incubation time: 10–60 minutes; linear detection range: 0.1–20 mM glucose uptake in cell-based assay |
| Sensitivity / LOD | Detection of insulin-stimulated glucose uptake increase of 1.5–3 fold over basal levels in adipocyte and muscle cell models |
| Specificity | 2-NBDG transport competes with D-glucose for GLUT-mediated uptake; no significant transport by sodium-dependent glucose transporters (SGLTs); fluorescent signal is specific to phosphorylated intracellular 2-NBDG after washing away extracellular dye |
| Reaction Conditions / Protocol | Serum-starve cells (2–6 hours) to reduce basal glucose uptake; wash with glucose-free buffer; treat with insulin or test compounds (15–30 min); add 2-NBDG in glucose-free buffer (100 μM final); incubate 10–60 min at 37°C; wash twice with cold PBS to remove extracellular dye; measure fluorescence in microplate reader or analyze by flow cytometry |
| Components / Formulation | 2-NBDG (lyophilized, 5 mg), DMSO for reconstitution, Glucose-free Assay Buffer, Insulin positive control (recombinant human insulin, 100× solution), Phloretin (GLUT inhibitor) negative control, detailed protocol for plate reader and flow cytometry formats |
| Storage Conditions | 2-NBDG lyophilized: -20°C protected from light and moisture for 12 months; reconstituted stock (10 mM in DMSO): -20°C desiccated for 1 month; other buffer components: 2–8°C |
| Shelf Life | 12 months from date of manufacture |
| Package Specifications | 200 assays (96-well), 1,000 assays |
| Product Form | Lyophilized fluorescent glucose analog with assay buffers and controls |
| Quality Control | Each lot tested for insulin-stimulated glucose uptake in 3T3-L1 adipocytes (≥2-fold induction); 2-NBDG purity ≥95% by HPLC; linearity of fluorescence response with cell number confirmed |
| Key Features | Direct fluorescence detection without secondary enzymatic reactions; live-cell compatible for time-course experiments; wide dynamic range from basal to maximally stimulated glucose uptake; insulin and inhibitor controls included for assay validation |
For research use only, not for clinical use.
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