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 HOME > Cell Biology > Cell signaling |
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NAD/NADH Assay Kit (Red) |
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코 드 |
: 15257 |
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단 위 |
: 400 assay |
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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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NAD/NADH Assay
Fluorimetric NAD/NADH Assay Kit
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™ NAD/NADH Assay Kit provides a convenient method for sensitive detection of NAD, NADH and their ratio. The enzymes in the system specifically recognize NAD/NADH in an enzyme cycling reaction. There is no need to purify NAD/NADH 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 NAD/NADH in solutions, in cell extracts.
Sensitive : The kit detect as low as 10 picomoles of NAD/NADH 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 : NAD/NADH cycling enzyme mixture 2 bottles (lyophilized powder)
Component B : NADH sensor buffer 1 bottle (20 mL)
Component C : NADH standard (FW: 709) 1 vial (142 μg)
Brief Summary
→ Prepare NAD/NADH reaction mixture (50 μL)
→ Add NADH standards or test samples (50 μL)
→ Incubate at room temperature for 15 min-2hr
→ Read fluorescence at Ex 540 nm/Em 590 nm
References
1. Ziegenhorn J, Senn M, Bucher T. (1976) Molar absorptivities of beta-NADH and beta-NADPH. Clin Chem, 22, 151.
2. Ikegami T, Kameyama E, Yamamoto SY, Minami Y, Yubisui T. (2007) Structure and Properties of the Recombinant
NADH-Cytochrome b(5) Reductase of Physarum polycephalum. Biosci Biotechnol Biochem.
3. Kimura N, Fukuwatari T, Sasaki R, Shibata K. (2006) Comparison of metabolic fates of nicotinamide, NAD+
and NADH administered orally and intraperitoneally; characterization of oral NADH. J Nutr Sci Vitaminol (Tokyo), 52,
142.
4. O'Donnell JM, Kudej RK, LaNoue KF, Vatner SF, Lewandowski ED. (2004) Limited transfer of cytosolic NADH into
mitochondria at high cardiac workload. Am J Physiol Heart Circ Physiol, 286, H2237. |
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Nicotinamide adenine dinucleotide
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Product Code |
Unit |
Price |
Availability |
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15257 |
400 assay |
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