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| Caspase / Apoptosis
Detection |
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High Throughput Screening
- Apo3HTS |
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Caspase Poly,1,2,3,7,8,9,10
Detection - FAM-FMK |
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Caspase Poly, 1,2,3,8,9,10,
Detection
- (SR-FMK) |
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Antibody Specific Caspase 3 Detection
- Apo Active 3 (FITC) |
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Antibody Specific Caspase 3 Detection
- Apo Active 3 (PE) |
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| NOS/ROS Detection Kits |
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| Fluorescent Enzymatic
Assays |
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| Mitochondria
Membrane Potential detection |
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| aCella - Bioluminescence
Assays Kits |
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| ELISA
Products |
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| Cathepsin B,K,L
Detection Kits |
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Fluoro NADP™ NEW!
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
#
|
Size
|
Price
(US$) |
| NADPH 100-2 |
100 Tests |
$495 |
|
| * Please call
888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for
volume pricing |
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| |
Product Data Sheet |
Protocol |
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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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