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Fluoro NADP
Fluorescent NADP/NADPH
Detection Kit - Detection of NADP/NADPH in
tissue extracts /cell samples
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| 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
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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.
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| Assay
Principle |
The Fluoro NADP/NADPH
detection kit utilizes a non-fluorescent
detection reagent, which is oxidized 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.
Reaction:
1. NADPH + non-fluorescent detection reagent +
electron coupler → fluorescent analog + NADP
2. NADP + enzyme coupled reaction → NADPH (then
proceeds to reaction 1).
Excitation:
530-570nm and Emission at 590-600nm
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References |
1.
Comprehensive Invited Review WEIHAI YING. NAD /NADH
and NADP /NADPH in Cellular Functions and Cell
Death: Regulation and Biological Consequences.
ANTIOXIDANTS &REDOX SIGNALING Volume 10,Number
2,2008.
2. Bedard K and Krause KH. The NOX family of ROS-generating
NADPH oxidases: physiology and
pathophysiology.Physiol Rev 87:245 –313,2007.
3. Lowry, Oliver H., Passonneau, Janet V. and
Rock, Martha K. The Stability of Pyridine
Nucleotides. The Journal of Biological
Chemistry, 236, #10, 1961. |
| Kit
contents (for 100 assays) |
1. Enzyme Mix
170μL Part 6022
.......................... 4-8C
2. NADP/NADPH Lysis Solution 12.5mL Part 3050
............................4-8C
3. NADP Extraction Buffer 22 mL Part
3046 (Green label) ..........4-8C
4. NADPH Extraction Buffer 22 mL Part
3047 (Magenta label)........4-8C
5. Reaction Buffer 11 mL.
Part 3049 ............................4-8C
6. Standard curve diluent 15 ml. Part
3052............................4-8C
7. NADPH Detection reagent 110 μL Part
4019...........................-20C
8. 3X Substrate Mix 6mL Part
3048........................... -20C
9. NADPH Standard- 3 vials dried Part
7014.............................-20C
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The following kits are available: |
Catalog
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Size
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Price (US$) |
| NADPH 100-2 |
100 Tests |
$495 |
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Comparison of NADPH & NADH Std Curves

Comparison of NADP & NADPH Std Curves
NADP and NADPH levels were assayed using the
Fluoro NADP/NADPH kit. Yield from 6 X105
Jurkat cells (n=3). |
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