Revision 2
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
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UniProt ID:

#P11413

Entrez-Gene Id:

2539

Product Information

Specificity / Sensitivity

The Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit detects sample G6PD activity. The presence of NADH and NADPH may interfere with the assay.

Species Reactivity:

All Species Expected

Product Description

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.

Background

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).

  1. Au, S.W. et al. (2000) Structure 8, 293-303.
  2. Jiang, P. et al. (2011) Nat Cell Biol 13, 310-6.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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Revision 2
#12581

Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit

Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit: Image 1 Expand Image
图 1. 葡萄糖-6-磷酸脱氢酶 (G6PD) 检测的示意图。在有 NADP 的情况下,葡萄糖-6-磷酸 (G6P) 被 G6PD 氧化,从而产生 6-磷酸葡萄糖酸内酯和 NADPH。所产生的 NADPH 随后被心肌黄酶循环系统扩增,从而产生强荧光试卤灵分子。
Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit: Image 2 Expand Image
图 2. 该检测系统可分别忽略每种检测组分,所产生的 RFU 与包含所有检测组分的对照检测的 RFU 进行对比。
Glucose-6-Phosphate Dehydrogenase (G6PD) Activity Assay Kit: Image 3 Expand Image
图 3. 未经处理和已经 G6PD 抑制剂 DHEA (0.5 mM) 处理的 Jurkat 细胞的裂解物蛋白浓度与相对荧光单位 (RFU) 之间的关系如图所示。如该图所示,G6PD 抑制剂 DHEA 能有效抑制这种链反应。