Figure 2. Each assay component is individually omitted from the assay system and the resultant RFU is compared to that of a control test that contains all of the assay components.
Figure 3. The relationship between the protein concentration of lysates from untreated and G6PD inhibitor DHEA (0.5 mM) treated Jurkat cells and relative fluorescence (RFU) is shown. The G6PD inhibitor DHEA can effectively inhibit this chain reaction as shown in this figure.
Figure 1. Schematic diagram of glucose-6-phosphate dehydrogenase (G6PD) assay. Glucose-6-phosphate (G6P) is oxidized by G6PD in the presence of NADP, which generates 6-phosphogluconolactone and NADPH. The generated NADPH is then amplified by the diaphorase-cycling system to produce highly fluorescent resorufin molecules.
|Product Includes||Quantity||Solution Color||Cap Color|
|Tris Assay Buffer||25 ml|
|G6PDH Substrate (40X)||250 µl||Blue|
|G6PDH Cofactor (100X)||100 µl||Yellow||Yellow|
|NADP+ (100X)||100 µl||White|
|G6PDH Developer (100X)||100 µl||Blue||Brown|
|G6PDH Positive Control (100X)||50 µl||Brown|
|PathScan® Sandwich ELISA Lysis Buffer (1X) 7018||30 ml|
The Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit contains the necessary reagents for rapid, sensitive, and simple detection of G6PD activity in various samples. In the assay, glucose-6-phosphate (G6P), in the presence of NADP, is oxidized by G6PD to generate 6-phosphogluconolactone and NADPH. The generated NADPH is then amplified by the diaphorase-cycling system to produce highly fluorescent resorufin molecules (see Figure 1). The relative fluorescent units (RFU) can then be determined using a plate reader with excitation about 540 nm and emission about 590 nm. The magnitude of RFU is proportional to G6PD activity in the sample.
The Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit detects sample G6PD activity. The presence of NADH and NADPH may interfere with the assay.
Glucose-6-phosphate dehydrogenase (G6PD) catalyses the first, and rate-limiting, step of the pentose phosphate pathway (1). The NADPH generated from this reaction is essential to protect cells from oxidative stress (1). Research studies have shown that p53 interacts with G6PD and inhibits its activity, therefore suppressing glucose consumption through the pentose phosphate pathway (2). In cancer cells with p53 mutations, the increased glucose consumption is directed towards increased biosynthesis, which is critical for cancer cell proliferation (2).
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