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HOME > Cell Biology > Cell signaling
Total NAD/NADH Assay Kit
   
코    드 : 15258
단     위 : 400 assays
공 급 원 : AAT Bioquest
가     격 : 문의
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  NAD/NADH Assay
Amplite™ Colorimetric Total 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


Key Features

Broad Application : Can be used for quantifying NAD/NADH in solutions, in cell extracts.
Sensitive : The kit detect as low as 30 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 absorbance at 575±5 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.
   
 
  15258

Product Code

Unit

Price

Availability

15258

400 assay