Description
Introduction
Nicotinamide adenine dinucleotide phosphate (NADP+) is utilized in anabolic reactions, comparable to lipid and nucleic acid synthesis, which require NADPH as a decreasing agent. NADPH is the diminished type of NADP+, and NADP+ is the oxidized type of NADPH. In cells, NADPH performs the function of a provider of decreasing energy and is primarily concerned in sustaining optimum redox metabolism. A simplified assay for the measurement of NAD and NADP is essential to understanding the roles of those pyridine nucleotides in regular and irregular cells.
NADPH is produced within the oxidative part of the pentose phosphate pathway in cells, a multifunctional pathway whose main function is to generate decreasing energy within the type of NADPH. NADPH is a cofactor for enzymes that synthesize energy-rich molecules and supply the decreasing equivalents for the oxidation-reduction concerned in defending the cell from the toxicity of reactive oxygen species (ROS) and NADPH oxidase-dependent ROS technology. Each NAD and NADP have been proven to affect hemoglobin affinity for oxygen in erythrocytes.
In-plant cells, NADPH is used because the decreasing energy for the biosynthetic reactions within the Calvin cycle of photosynthesis (1-2).
Cell Expertise’s Fluoro NADP/NADPH offers a extremely dependable, delicate fluorometric assay for the quantification of NADP, NADPH and their ratio in organic samples.
Key Advantages
- Detection of NADP/NADPH content material in cells or tissue extracts.
- Detection of NADP/NADPH ranges in antioxidation and oxidative stress.
- Detection of NADP/NADPH ranges in cell dying, vitality metabolism, and mitochondria perform.
- Species Impartial – NADP/NADPH detection in Bacterial, fungal and plant cells.
- Extremely Delicate – Detects as much as 4nM NADP and NADPH.
- Extremely Particular – No Cross reactivity with NAD/NADH.
- Straightforward to Use – 96 effectively Fluorescent Plate reader readout.
Summary
NAD is a key determinant of mobile vitality metabolism. In distinction, its phosphorylated kind, NADP, performs a central function in biosynthetic pathways and antioxidant defence. The diminished types of each pyridine nucleotides are fluorescent in residing cells however they can’t be distinguished, as they’re spectrally similar. Right here, utilizing genetic and pharmacological approaches to perturb NAD(P)H metabolism, we discover that fluorescence lifetime imaging (FLIM) differentiates quantitatively between the 2 cofactors.
Systematic manipulations to alter the stability between oxidative and glycolytic metabolism counsel that these states don’t instantly impression NAD(P)H fluorescence decay charges. The lifetime modifications noticed in cancers thus doubtless mirror shifts within the NADPH/NADH stability. Utilizing a mathematical mannequin, we use these experimental knowledge to quantify the relative ranges of NADH and NADPH in several cell forms of a fancy tissue, the mammalian cochlea. This reveals NADPH-enriched populations of cells, elevating questions on their distinct metabolic roles.

Introduction
The mobile redox state is the central regulator of vitality manufacturing and middleman metabolism, taking part in an important function in well being and illness. The nicotinamide adenine dinucleotide (NAD+/NADH) and nicotinamide adenine dinucleotide phosphate (NADP+/NADPH) redox {couples} are the main determinants of redox state within the cell. Nevertheless, these have interaction in distinct metabolic pathways. NAD drives ATP manufacturing within the cytosol by glycolysis and within the mitochondria by oxidative phosphorylation, whereas the phosphorylated analogue NADP governs lipid, amino acid and nucleotide biosynthetic pathways and the defence towards reactive oxygen species by glutathione (GSH). Free radical technology is due to this fact decided by the redox state of NAD, whereas NADP redox state is vital to antioxidant defence3. The relative abundance of the 2 pyridine nucleotides and their redox stability thus mediates cell destiny in a variety of ailments, together with most cancers, diabetes and neurodegeneration. Quantifying their behaviour is due to this fact important in understanding the function of metabolism in these ailments. Nevertheless, separating the contributions of the 2 swimming pools in intact tissues has confirmed technically difficult4.
Within the 1960s, Britton Probability et al.5 confirmed that stay tissues illuminated with ultraviolet gentle emit blue fluorescence, arising primarily from mitochondrial NADH. The nicotinamide moiety of NADH absorbs gentle of wavelength 340±30 nm and emits fluorescence at 460±50 nm. As NADP is phosphorylated at a distant web site of the molecule, the fluorescence properties of the nicotinamide ring of NADPH are similar to these of NADH6,7. Thus, modifications in autofluorescence depth could mirror modifications in both [NADH] or [NADPH], usually denoted as NAD(P)H to point the unsure origin of the sign.
fluoro-Dapagliflozin |
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C5119-5 | ApexBio | 5 mg | EUR 238.8 |
Description: Ki: 5.3 and 330 nM for SGLT2 and SGLT1, respectivelyfluoro-Dapagliflozin is a selective sodium-glucose cotransporter (SGLT) inhibitor.Human Na(+)-D-glucose cotransporter (hSGLT) inhibitors constitute the newest class of diabetes drugs, blocking up to in-vivo 50% of renal glucose reabsorption. |
fluoro-Dapagliflozin |
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C5119-50 | ApexBio | 50 mg | EUR 1363.2 |
Description: Ki: 5.3 and 330 nM for SGLT2 and SGLT1, respectivelyfluoro-Dapagliflozin is a selective sodium-glucose cotransporter (SGLT) inhibitor.Human Na(+)-D-glucose cotransporter (hSGLT) inhibitors constitute the newest class of diabetes drugs, blocking up to in-vivo 50% of renal glucose reabsorption. |
Active NADP-dependent Isopropanol Dehydrogenase (ADH1-NADP) |
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7605-1000 | Biovision | EUR 444 |
Active NADP-dependent Isopropanol Dehydrogenase (ADH1-NADP) |
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7605-200 | Biovision | EUR 183.6 |
NADP, disodium salt |
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2736-100 | Biovision | EUR 183.6 |
NADP, disodium salt |
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2736-1000 | Biovision | EUR 940.8 |
NADP, disodium salt |
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2736-500 | Biovision | EUR 548.4 |
NADP, sodium salt |
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2737-100 | Biovision | EUR 183.6 |
NADP, sodium salt |
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2737-1000 | Biovision | EUR 940.8 |
NADP, sodium salt |
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2737-500 | Biovision | EUR 548.4 |
NADP (sodium salt) |
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HY-F0002 | MedChemExpress | 500mg | EUR 325.2 |
NADP (disodium salt) |
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HY-F0002A | MedChemExpress | 500mg | EUR 325.2 |
5-fluoro 203 |
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C4114-10 | ApexBio | 10 mg | EUR 289.2 |
Description: 5-fluoro 203 (5F-203) is an antitumor agent and cytotoxic compound that acts as a potent AhR agonist [1][2][3]. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor involved in regulating xenobiotic-metabolizing enzymes such as cytochrome P450. |
5-fluoro 203 |
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C4114-25 | ApexBio | 25 mg | EUR 582 |
Description: 5-fluoro 203 (5F-203) is an antitumor agent and cytotoxic compound that acts as a potent AhR agonist [1][2][3]. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor involved in regulating xenobiotic-metabolizing enzymes such as cytochrome P450. |
5-fluoro 203 |
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C4114-5 | ApexBio | 5 mg | EUR 184.8 |
Description: 5-fluoro 203 (5F-203) is an antitumor agent and cytotoxic compound that acts as a potent AhR agonist [1][2][3]. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor involved in regulating xenobiotic-metabolizing enzymes such as cytochrome P450. |
Benzeneethanamine,2-fluoro- |
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abx181242-50g | Abbexa | 50 g | EUR 510 |
Pyrazine, 2-fluoro- |
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20-abx182629 | Abbexa |
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5-Fluoro-tryptaminehydrochloride |
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20-abx185281 | Abbexa |
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NADP / NADPH Assay Kit |
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abx096008-100Assays | Abbexa | 100 Assays | EUR 566.4 |
NADP+/NADPH Assay Kit |
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MET-5018 | Cell Biolabs | 100 assays | EUR 686.4 |
Description: Our NADP+/NADPH Assay Kit detects NADP+ and NADPH in cell and tissue lysates. Total NADP+/NADPH can be detected or samples can be treated with an acid or base treatment to specifically detect NADP+ or NADPH. This assay uses an enzymatic cycling reaction that reduces NADP+ to NADPH, which then reacts with a colorimetric probe and is detected with a spectrophotometric plate reader at 450nm. NADP+/ NADPH levels in unknown samples are calculated based on the provided NADP+ standard curve. |
NADP/NADPH Assay Kit |
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Z5030038 | Biochain | 100 assays | EUR 966 |
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol. |
6-fluoro-DL-Tryptophan |
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C5214-100 | ApexBio | 100 mg | EUR 164.4 |
Description: 6-fluoro-DL-Tryptophan is a serotonin (5-HT) synthesis inhibitor.Serotonin or 5-hydroxytryptamine (5-HT), a monoamine neurotransmitter, is biochemically derived from tryptophan. |
6-fluoro-DL-Tryptophan |
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C5214-250 | ApexBio | 250 mg | EUR 265.2 |
Description: 6-fluoro-DL-Tryptophan is a serotonin (5-HT) synthesis inhibitor.Serotonin or 5-hydroxytryptamine (5-HT), a monoamine neurotransmitter, is biochemically derived from tryptophan. |
6-fluoro-DL-Tryptophan |
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C5214-500 | ApexBio | 500 mg | EUR 374.4 |
Description: 6-fluoro-DL-Tryptophan is a serotonin (5-HT) synthesis inhibitor.Serotonin or 5-hydroxytryptamine (5-HT), a monoamine neurotransmitter, is biochemically derived from tryptophan. |
2-fluoro Palmitic Acid |
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C3924-10 | ApexBio | 10 mg | EUR 182.4 |
Description: 2-fluoropalmitic acid inhibits sphingosine biosynthesis and long-chain acyl-CoA synthetase [1]. The length of the carbon chain of the fatty acid species defines the substrate specificity for the different acyl-CoA synthetases (ACS). |
2-fluoro Palmitic Acid |
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C3924-25 | ApexBio | 25 mg | EUR 336 |
Description: 2-fluoropalmitic acid inhibits sphingosine biosynthesis and long-chain acyl-CoA synthetase [1]. The length of the carbon chain of the fatty acid species defines the substrate specificity for the different acyl-CoA synthetases (ACS). |
2-fluoro Palmitic Acid |
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C3924-50 | ApexBio | 50 mg | EUR 582 |
Description: 2-fluoropalmitic acid inhibits sphingosine biosynthesis and long-chain acyl-CoA synthetase [1]. The length of the carbon chain of the fatty acid species defines the substrate specificity for the different acyl-CoA synthetases (ACS). |
4-Fluoro-2-phenylbenzoicacid |
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20-abx180542 | Abbexa |
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