QuantiChrom™ G6PDH Inhibitor Screening Kit

Protocol SDS

Application

• For evaluation and high-throughput screen (HTS) of glucose-6-phosphate dehydrogenase modulators.

Key Features

• Safe. Non-radioactive assay.
• High-throughput. Homogenous “mix-incubate-measure” type assay. Can be readily automated on HTS liquid handling system.
• Rapid and reliable. Can be completed in 30 minutes and no 37°C heater is needed.

Method

• OD565nm

Samples

• Compounds that affect glucose-6-phosphate dehydrogenase activity

Species

• All

Procedure

• 30 min

Size

• 100 tests

Shelf Life

• 6 months

More Details

GLUCOSE-6-PHOSPHATE DEHYDROGENASE (G6PDH) is a cytosolic enzyme in the pentose phosphate pathway which supplies reducing energy to cells by maintaining the level of the co-enzyme nicotinamide adenine dinucleotide phosphate (NADPH). G6PDH reduces nicotinamide adenine dinucleotide phosphate (NADP) to NADPH while oxidizing glucose-6-phosphate (G6P). Humans with a genetic deficiency of G6PDH are predisposed to non-immune hemolytic anemia. Recently, studies have found G6PDH plays a critical role in the survival, proliferation, and metastasis of cancer cells. Therefore, inhibitors of the enzyme are attractive candidates for new cancer therapeutics. BioAssay System’s non-radioactive, colorimetric G6PDH assay is based on the reduction of the tetrazolium salt MTT in a NADPH-coupled enzymatic reaction to a reduced form of MTT which exhibits an absorption maximum at 565 nm. The increase in absorbance at 565 nm is proportional to the enzyme activity. The percent inhibition of a test compound can be determined by comparing the activity of G6PDH treated with a test compound to the activity of untreated G6PDH.

Does the kit come with the G6PDH enzymes or control inhibitors?

No, these are not included.

How do I store the kit?

Store all components at -20°C for long term storage.

What species of G6PDH was the kit optimized for?

The kit was optimized for L. Mesenteroides G6PDH.

What if I am interested in testing inhibition of a species besides L. Mesenteroides?

We recommend that you experimentally determine the K_m and then adjust the volume of substrate so that the final concentration of the substrate in the 100 µL Reaction Volume is near the K_m.

What G6PDH inhibitor was tested using this kit?

Zn(NO₃)₂ in dH₂0.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.