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2-NBDG Glucose Uptake Assay Kit (Cell-Based)

Non-radioactive method to detect and accurately measure glucose uptake via FACS and microscopy
Catalog #: K682
$405.00

Product Details

Cat # +Size K682-50
Size 50 assays
Detection Method FACS (488 nm excitation laser) and fluorescent microscope (excitation range 420 nm-495 nm).
Applications Measurement of glucose uptake in response to insulin, growth factors, cytokines, mitogens and nutrients, etc.
Dual-staining of glucose transporters and glucose uptake
Analysis of glucose metabolism and cell signaling in various cell types
Screening of anti-diabetic compounds
Features & Benefits • Easy-to-use
• Non-radioactive
• Image and accurately measure glucose uptake in cultured cells in response to insulin, growth factors etc.
Kit Components • Analysis Buffer (50X)
• 2-NBDG Reagent (100X)
• Glucose Uptake Enhancer
• Phloretin (100X)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Glucose is a ubiquitous energy source in most organisms and plays a pivotal role in cellular metabolisms and homeostasis. Cancer cells exhibit increased glucose uptake to support their high proliferation rate. 2-NBDG (2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]-D-glucose) 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. 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.


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What is principle of the assay and can we use your KIT without glucose uptake enhancer, replacing it with insulin or other growth factors?
2-NBDG (2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl) amino]-Dglucose) is a fluorescent deoxyglucose analog that can be taken up by cells through glucose transporters, but it is not metabolized by cells and gets accumulated inside the cells. You can use this kit to look at the influence of insulin and other growth factors. You can use the kit with and without the glucose uptake enhancer.
Can the probe be used for animal experiments?
The fluorescent molecule has an excitation at 494nm, em 551nm. Typically this is not a suitable wavelength for in vivo animal imaging. For animal imaging applications its preferable to choose more red-shifted probes that have emission wavelengths >=650nm. However, I am not sure what kind of animal applications she is interested in- for tissue based studies this is probably ok or for superficial models like mouse xenografts. We don’t have a formulation designed for animal studies.
Can we use frozen samples with this assay?
We would recommend using fresh samples. However, frozen samples can be used too provided they have not undergone multiple freeze thaw cycles.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6-12 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Can FBS be completely avoided during treatment of cells with Test Compounds?
It is not recommended to avoid FBS completely as it will keep the cells healthy during the assay process. Also, our assay buffer has FBS in it. Cell death can occur rapidly in the complete absence of serum and you would need a minimum amount of serum to keep the cells happy.
Can a plate reader be used for the fluorometric detection?
No, you cannot use a plate reader for this assay as you are using live cells and not lysates. The detection method is flow cytometry or fluorescence microscopy.
Lynnea R. Waters, Ampk regulates IgD expression but not energy stress with B cell activation. Sci Rep, June 2019;  31160601.
Man-Hsin Hung, Canagliflozin inhibits growth of hepatocellular carcinoma via blocking glucose-influx-induced β-catenin activation. Cell Death Dis, May 2019; 31142735.
For more citations of this product click here