NADP/NADPH Assay Kit (Fluorometric) (BA0107)

NADP/NADPH Assay Kit

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 redox state of cells or tissue. Simple, direct and automation-ready procedures for measuring NADP⁺/NADPH concentration are very desirable.

Assay Genie's 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 Ex/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.

Applications

For quantitative determination of NADP and NADPH and evaluation of drug effects on their metabolism.

Key Features

• Sensitive and accurate. Detection limit of 0.01 uM and linearity up to 1 uM 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.

NADP/NADPH Assay Kit Information:

Kit Includes:	Assay Buffer: 10 mL Enzyme A: 120 mL Glucose (1 M): 1.5 mL Enzyme B: 120 mL Probe: 750 mL NADP Standard: 0.5 mL NADP/NADPH Extraction Buffers: each 12 mL
Kit Requires:	Pipetting (multi-channel) devices. Black, flat bottom 96-well plates and fluorescent plate reader capable of reading at FL »ex/em = 530/585 nm.
Method of Detection:	FL530/585nm
Detection Limit:	0.01 uM
Samples:	Cell, tissue extracts etc
Species:	All
Protocol Length:	30 min
Size:	100 tests
Storage:	Store all reagents at -20°C
Shelf Life:	6 months