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 HOME > Cell Biology > Cell signaling |
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NADP/NADPH Assay Kit (Red) |
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코 드 |
: 15259 |
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단 위 |
: 400 assays |
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공 급 원 |
: AAT Bioquest |
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가 격 |
: 문의 |
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다운로드 |
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NADP_NADPH Assay
Amplite™ Fluorimetric NADP/NADPH Assay Kit *Red Fluorescence*
Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. NADH is the reduced form of NAD+, and NAD+ is the oxidized form of NADH. It forms NADP with the addition of a phosphate group to the 2' position of the adenyl nucleotide through an ester linkage. NADP is used in anabolic biological reactions, such as fatty acid and nucleic acid synthesis, which require NADPH as a reducing agent. In chloroplasts, NADP is an oxidizing agent important in the preliminary reactions of photosynthesis. The NADPH produced by photosynthesis is then used as reducing power for the biosynthetic reactions in the Calvin cycle of photosynthesis. The traditional NAD/NADH and NADP/NADPH assays are done by monitoring of NADH or NADPH absorption at 340 nm. This method suffers low sensitivity and high interference since the assay is done in the UV range that requires expensive quartz microplate. This Amplite™ NADP/NADPH Assay Kit provides a convenient method for sensitive detection of NADP, NADPH and their ratio. The enzymes in the system specifically recognize NADP/NADPH in an enzyme cycling reaction. There is no need to purify NADP/NADPH from sample mix. The enzyme cycling reaction significantly increases detection sensitivity. In addition, this assay has very low background since it is run in the red visible range that significantly reduces the interference from biological samples. The assay has demonstrated high sensitivity and low interference with 570 nm excitation 590 nm emission.
Key Features
Broad Application: Can be used for quantifying NADP/NADPH in solutions, in cell extracts.
Sensitive: The kit detect as low as 1 picomoles of NADP/NADPH in solution.
Continuous: Easily adapted to automation with no separation required.
Convenient: Formulated to have minimal hands-on time. No wash is required.
Non-Radioactive: No special requirements for waste treatment.
Kit Components
Component A: NADP/NADPH recycling enzyme mixture 2 bottles (lyophilized powder)
Component B: NADPH sensor buffer 1 bottle (20 mL)
Component C: NADPH standard (FW: 833.36) 1 vial (167 μg)
Brief Summary
→ Prepare NADP/NADPH reaction mixture (50 μL)
→ Add NADPH standards or test samples (50 μL)
→ Incubate at room temperature for 15 min-2hr
→ Read fluorescence at Ex 570 nm/Em 590
References
1. Hedeskov CJ, Capito K, Thams P. (1987) Cytosolic ratios of free [NADPH]/[NADP+] and [NADH]/[NAD+] in mouse pancreatic islets, and nutrient-induced insulin secretion. Biochem J, 241,161.
2. Gaetani GF, Ferraris AM, Sanna P, Kirkman HN. (2005) A novel NADPH:(bound) NADP+ reductase and
NADH:(bound) NADP+ transhydrogenase function in bovine liver catalase. Biochem J, 385, 763.
3. Kobayashi K, Miura S, Miki M, Ichikawa Y, Tagawa S. (1995) Interaction of NADPH-adrenodoxin reductase with NADP+ as studied by pulse radiolysis. Biochemistry, 34, 12932.
4. Marino D, Gonzalez EM, Frendo P, Puppo A, Arrese-Igor C. (2006) NADPH recycling systems in oxidative stressed pea nodules: a key role for the NADP(+)-dependent isocitrate dehydrogenase. Planta. |
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15259
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Product Code |
Unit |
Price |
Availability |
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15259 |
400 assay |
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