2-Deoxyglucose (2DG) Uptake Measurement Kit - Cosmo Bio Co.,Ltd.

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2-Deoxyglucose (2DG) Uptake Measurement Kit

Catalog No.: CSR-OKP-PMG-K01TE
Size: 1KIT
[25 tests]
Price: ¥45000
$600
catalog info: Catalog 2012-p133
Storage: -20C
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Purpose: Direct measurement of 2DG6P without the use of radioisotope.
component: Solution A: 1,800 ul x 1 tube
Solution B (Acid solution): 500 ul x 1 tube
Solution C (Acid neutralizing solution): 500 ul x 1 tube
Solution D: 850 ul x 1 tube
Solution E (Alkali solution): 500 ul x 1 tube
Solution F (Alkali Neutralizing solution): 500 ul x 1 tube
Solution G: 1,100 ul x 1 tube
1 mM 2DG6P solution: 250 ul x 1 tube
Sample diluent buffer Concentrate (100-fold concentrated solution): 1.5 ml x 1 tube
Substrate buffer: 6.5 ml x 1 vial
DTNB Substrate (powder): 3 vials
Low G6PDH: 13 ul x 1 tube
High G6PDH: 130 ul x 1 tube
GR: 10 ul x 1 tube

Other:

 Background 
Measurement of 2-deoxyglucose (2DG) uptake in tissues and cells is a reliable approach with which to estimate glucose uptake and thereby to explore the regulation of glucose metabolism and mechanism of insulin resistance. Radioisotope-labeled 2DG is usually used for the measurement of 2DG uptake both in vivo and in vitro. However the radioisotope (RI) method is required a specialized facility for RI in strict limitation and cannot be handled in ordinal laboratories. Furthermore, radioactive 2DG administered into cultured cells remains in the extracellular space, and therefore the results must be corrected by separating the extracellular 2DG and intracellular 2DG/2DG-6-phosphate (2DG6P) in the cells.
Thiskit is based on theenzymatic method for the direct measurement of 2DG6P amount without any use of radioisotope materials (Saito K and Minokoshi Y, et al. Analytical Biochem 412: 9-17, 2011).
 Advantages 
1. No RI materials are required, and 2DG uptake can be measured in any ordinal laboratories.
2. Direct measurement of 2DG6P amount accumulated in target cells.
3. High sensitivity with the use of enzyme-recycling amplification reaction.




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A recycling enzymatic amplification system measures NADPH produced by the in vitro oxidation of 2DG6P accumulated in cells following 2DG uptake. 1) So as not to effect glucose metabolism, only a small amount of 2DG is added to live cells. Incorporated 2DG is converted by cell metabolism to 2D6GP, which accumulates in cells. Cell lysates are then prepared. 2) To eliminate detection of G6P, G6P is oxidized (to 6PG) with NAD+ and a low concentration of G6PDH. 3) 2DG6P levels are quantitated by measuring the amount of NADPH produced during 2DG6P oxidation (with NADP+ and a high concentration of G6PDH) in a phtometric recycling amplification/detection system All reaction steps are conveniently performed in a single well by the sequential addition of premixed reagents. Ideal for assay automation

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Comparison between this kit and RI method

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Temporal change of O.D. for different concentrations of 2DG6P

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Experimental example 1 - 2-deoxyglucose (2DG) uptake by 3T3-L1 cells

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Experimental Example 2 - 2-deoxyglucose (2DG) uptake by human adipocytes

Reference:

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