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CheKine™ Coenzyme Ⅱ NADP(H) Assay Kit

CheKine™ Coenzyme Ⅱ NADP(H) Assay Kit

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Specification

Product name CheKine™ Coenzyme Ⅱ NADP(H) Assay Kit
Applications notes Abbkine CheKine™ Coenzyme Ⅱ NADP(H) Assay Kit provides a convenient tool for sensitive detection of the NADP and NADPH, and their ratio in various tissues and subcellular organelles. This Kit is based on enzyme cycling reaction (It does not recognize NAD+/NADH), in which the formed NADPH reduces a formazan (MTT) reagent. The intensity of the reduced product color, measured at OD565 nm, is proportionate to the NADP+ /NADPH concentration in the sample.

Product Properties

Kit components • Assay Buffer
• Glucose (1 M)
• WST-8 Solution
• Enhancer Solution
• NADP Cycling Enzyme Mix
• NADPH standard (10 mM)
• NADP Extraction Buffer
• NADPH Extraction Buffer
Features & Benefits • Detect NADP+/NADPH concentrations and ratios in cell or tissue extracts.
• Detection limit of 0.1 µM and linearity up to 4 µM NADP+/NADPH in 96-well plate assay.
Calibration range 0.5 µM-10 µM
Usage notes • Addition of Working Reagent should be quick and mixing should be brief but thorough.
• The following substances interfere and should be avoided in sample preparation. EDTA (>0.5 mM), ascorbic acid, SDS (>0.2%), sodium azide, NP-40 (>1%) and Tween-20 (>1%).
Storage instructions Storage at –20°C and Keep from light immediately upon receipt. Kit has a storage time of 6 months from receipt. Refer to list of materials supplied for storage conditions of individual components.
Shipping Gel pack with blue ice.
Precautions The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.

Additional Information

Background Nicotinamide adenine dinucleotide phosphate (NADP) is an enzymatic cofactor involved in many redox reactions where it cycles between the reduced (NADPH) and oxidized (NADP) forms. NADP is also involved in biosynthetic reactions such as lipid and nucleic acid synthesis where it functions as a reducing agent. The oxidative branch of the pentose phosphate pathway (PPP) is the major source of NADPH produced in animal cells. 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 tissues.

Image & description

Fig. Standard Curve of NADPH in 96-well plate assay. The y-axis is ΔOD and the x-axis is NADPH concentration (uM).

Fig. Standard Curve of NADPH in 96-well plate assay. The y-axis is ΔOD and the x-axis is NADPH concentration (uM).

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