EnzyFluo™ NADP/NADPH Assay Kit

Protocol SDS

Application

• For quantitative determination of NADP and NADPH and evaluation of drug effects on their metabolism (This product is a replacement for the discontinued product EFNP-100).

Key Features

• Sensitive and accurate. The detection limit of 0.01 µM and linearity up to 1 µM NADP+/NADPH in 96-well plate assay.
• Convenient. The procedure involves adding a single working reagent and reading the fluorescence at time zero and 30 min. Room temperature assay.
• High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method

• FL530/585nm

Samples

• Cell, tissue extracts, etc

Species

• All

Procedure

• 30 min

Size

• 100 tests

Detection Limit

• 0.01 µM

Shelf Life

• 6 months

More Details

Pyridine nucleotides play an important role in metabolism and, thus, there is continual interest in monitoring their concentration levels. Quantitative determination of NADP⁺/NADPH has applications in research pertaining to energy transformation and the redox state of cells or tissue. Simple, direct, and automation-ready procedures for measuring NADP⁺/NADPH concentration are very desirable. BioAssay Systems EnzyFluo™ NADP⁺/NADPH assay kit is based on a glucose dehydrogenase cycling reaction, in which the formed NADPH reduces a probe into a highly fluorescent product. The fluorescence intensity of this product, measured at λex/em = 530/585 nm, is proportional to the NADP⁺/NADPH concentration in the sample. This assay is highly specific for NADP⁺/NADPH and with minimal interference (<1%) by NAD⁺/NADH. Our assay is a convenient method to measure NADP⁺, NADPH, and their ratio.

When tissue is used, should it be freshly obtained? Or is -80°C storage ok?

If direct sample processing is not possible, we recommend snap freezing tissue samples in liquid nitrogen and keeping them either in liquid nitrogen or at -80°C until further processing.

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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.