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 HOME > Cell Biology > Cell signaling |
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NAD/NADH Ratio Assay Kit |
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코 드 |
: 15263 |
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단 위 |
: 250 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 Ratio Assay Kit
Amplite™ Fluorimetric NAD/NADH Ratio 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. 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. Our Amplite™ NAD/NADH Ratio 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 Ex/Em = 540/590 nm. This Amplite™Fluorimetric NAD/NADH Assay Kit can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation with no separation steps required.
Key Features
Broad Application : Can be used for quantifying NAD/NADH in solutions and in cell extracts.
Sensitive : Detect as low as 10 picomoles of NAD/NADH in solution.
Continuous : Easily adapted to automation without a separation step.
Convenient : Formulated to have minimal hands-on time.
Non-Radioactive : No special requirements for waste treatment.
Kit Components
Component A : NAD/NADH Recycling Enzyme Mixture 2 bottles (lyophilized powder)
Component B : NADH Sensor Buffer 1 bottle (20 mL)
Component C : NADH Standard 1 vial (142 μg)
Component D : NADH Extraction Solution 1 bottle (10 mL)
Component E : NAD Extraction Solution 1 bottle (10 mL)
Component F : NAD/NADH Control Solution 1 bottle (10 mL)
Component G : NAD/NADH Lysis Buffer 1 bottle (10 mL)
Brief Summary
→ Prepare 25 μL of NADH standards and/or test samples
→ Add 25 μL of NADH or NAD Extraction Solution
→ Incubate at room temperature for 15 minutes
→ Add 25 μL of NAD or NADH Extraction Solution
→ Add 75 μL of NAD/NADH reaction mixture
→ Incubate at RT for 15 minutes to 2 hours
→ Monitor fluorescence intensity at Ex/Em = 540/590 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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15263
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Product Code |
Unit |
Price |
Availability |
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15263 |
250 assay |
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