Fluoro NADP

$520.00$2,495.00

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Introduction

Nicotinamide adenine dinucleotide phosphate (NADP+) is used in anabolic reactions, such as lipid and nucleic acid synthesis, which require NADPH as a reducing agent. NADPH is the reduced form of NADP+, and NADP+ is the oxidized form of NADPH. In cells, NADPH plays the role of a carrier of reducing power and is primarily involved in maintaining optimal redox metabolism. A simplified assay for the measurement of NAD and NADP is critical to understanding the roles of these pyridine nucleotides in normal and abnormal cells.

NADPH is produced in the oxidative phase of the pentose phosphate pathway in cells, a multifunctional pathway whose primary purpose is to generate reducing power in the form of NADPH. NADPH is a cofactor for enzymes that synthesize energy-rich molecules and provide the reducing equivalents for the oxidation-reduction involved in protecting the cell from the toxicity of reactive oxygen species (ROS) and NADPH oxidase-dependent ROS generation. Both NAD and NADP have been shown to influence hemoglobin affinity for oxygen in erythrocytes. In plant cells, NADPH is used as the reducing power for the biosynthetic reactions in the Calvin cycle of photosynthesis (1-2).

Cell Technology’s Fluoro NADP/NADPH provides a highly reliable, sensitive fluorometric assay for the quantification of NADP, NADPH and their ratio in biological samples.

Key Benefits

  • Detection of NADP/NADPH content in cells or tissue extracts.
  • Detection of NADP/NADPH levels in antioxidation and oxidative stress.
  • Detection of NADP/NADPH levels in cell death, energy metabolism, and mitochondria function.
  • Species Independent – NADP/NADPH detection in Bacterial, fungal and plant cells.
  • Highly Sensitive – Detects up to 4nM NADP and NADPH.
  • Highly Specific – No Cross reactivity with NAD/NADH.
  • Easy to Use – 96 well Fluorescent Plate reader readout.

Additional information

Kit Size

100, 500

The Fluoro NADP/NADPH detection kit utilizes a non-fluorescent detection reagent, which is reduced in the presence NADPH to produce its fluorescent analog and NADP. NADP is further converted to NADPH via an enzyme-coupled reaction. The enzyme reaction specifically reacts with NADP/NADPH and not with NAD/NADH.

fluorescent_nadp_nadph_1

Figure 1. Comparison of NADPH & NADH Std Curves

fluorescent_nadp_nadph_2

Figure 2. Comparison of NADP & NADPH Std Curves

Part#ReagentTemperature
Part # 6022Enzyme Mix, 170μL2-8C
Part # 3050NADP/NADPH Lysis Solution, 12.5mL2-8C
Part # 3057 (Green label)NADP Extraction Buffer, 22 mL2-8C
Part # 3058 (Magenta label)NADPH Extraction Buffer, 22 mL2-8C
Part # 3049Reaction Buffer, 11 mL.2-8C
Part # 3052Standard curve diluent, 15 ml.2-8C
Part # 4019NADPH Detection reagent, 110 μL-20C
Part # 30483X Substrate Mix, 6mL-20C
Part # 7014NADPH Standard, 3 vials dried-20C

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