Description
Key facts
Detection method:Fluorescent,
Sample types:Urine, Plasma, Serum, Other biological fluids, Whole Blood, Cell Lysate,
Assay type:Enzyme activity (quantitative),
Sensitivity:>= 0.3 mIU/mL,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
Glucose 6 Phosphate Dehydrogenase Activity Assay Kit (Fluorometric) (ab176722) provides a simple method for detecting glucose 6 phosphate dehydrogenase (G6PD) activity in a variety of biological samples such as serum, plasma and urine, as well as cell extracts.
G6PD assay principle
In the assay, G6PD present in the sample converts NADP
to NADPH, which can be specifically monitored by a fluorogenic NADPH sensor to yield a highly red fluorescence product. The signal can be easily detected at Ex/Em = 540/590 nm in a fluorescent microplate reader. The fluorescence intensity is proportional to the G6PD activity present in the sample.
The assay can detect glucose 6 phosphate dehydrogenase activity from as low as 0.3 mU/mL in a 100 μL reaction volume.
Other notes
Glucose 6 Phosphate Dehydrogenase (G6PD, G6PDH, EC 1.1.1.49) catalyzes the conversion of glucose-6-phosphate to 6-phosphoglucono-Δ-lactone, the first and rate-limiting step in the pentose phosphate pathway (PPP). It is a critical metabolic pathway that supplies reducing energy to cells (such as erythrocytes) by maintaining the level of co-enzyme nicotinamide adenine dinucleotide phosphate (NADPH). The production of NADPH is of great importance for tissues actively engaged in biosynthesis of fatty acids and/or isoprenoids, such as the liver, mammary glands, adipose tissue, and the adrenal glands. The NADPH also maintains the level of glutathione in these cells that helps protect the red blood cells against oxidative damage. Deficiencies in G6PD predispose individuals to non-immune hemolytic anemia.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Glucose 6 Phosphate Dehydrogenase also known as G6PD or G6P dehydrogenase plays an important role in the pentose phosphate pathway catalyzing the conversion of glucose 6-phosphate to 6-phosphoglucono-Δ-lactone while producing NADPH from NADP+. G6PD with a molecular mass of approximately 59 kDa is expressed in various tissues with high levels found in red blood cells liver and adrenal glands. This enzyme is vital for protecting cells from oxidative damage by supplying reductive capacity.
Biological function summary
Glucose 6-phosphate dehydrogenase is essential for maintaining cellular redox balance especially in cells lacking mitochondria like red blood cells. It functions as part of a monomer which can dimerize depending on the cellular needs and conditions. The activity of G6PD directly affects the production of NADPH which is necessary for the biosynthesis of nucleic acids and lipids and for maintaining reduced glutathione levels therefore sustaining the antioxidant capacity of the cell.
Pathways
Glucose 6-phosphate dehydrogenase is a pivotal enzyme in the oxidative phase of the pentose phosphate pathway which provides ribose 5-phosphate for nucleotide synthesis and NADPH for reductive biosynthetic reactions. G6PD connects with other enzymes in the pathway such as 6-phosphogluconate dehydrogenase contributing to the regulation of cellular metabolic needs. Its function is tightly linked to glucose metabolism and indirectly influences glycolytic processes.
Alterations in glucose 6-phosphate dehydrogenase activity lead to conditions such as G6PD deficiency which can cause hemolytic anemia when individuals are exposed to certain drugs infections or foods. This enzyme's deficiency is one of the most common enzymatic disorders in humans and is triggered by genetic mutations affecting G6PD function. Moreover G6PD has implications in cancer biology as its activity influences the proliferation and survival of cancer cells alongside other proteins like TP53 and NF-κB which are involved in cellular stress responses.
