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HOME > Cell Biology > Cell signaling
NADPH Assay Kit
   
코    드 : 15262
단     위 : 400 assay
공 급 원 : AAT Bioquest
가     격 : 문의
다운로드

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  NADH Assay
Amplite™ Fluorimetric 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™ NADPH Assay Kit provides a convenient method for sensitive detection of NADPH. The enzymes in the system specifically recognize NADPH in an enzyme cycling reaction. The enzyme cycling reaction significantly increases detection sensitivity.


Key Features

Broad Application : Can be used for quantifying NADPH in solutions, in cell extracts.
Sensitive : The kit detect as low as 1 nanomoles of 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 : NADPH recycling enzyme mixture          2 bottles (lyophilized powder)
Component B : NADPH assay buffer                              1 bottle (20 mL)
Component C : NADPH standard (FW: 833.36)                 1 vial (167 μg)

Brief Summary

→ Prepare 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 540 nm/Em 590 nm

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-adrenodoxinreductase 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 inoxidative stressed
   pea nodules: a key role for the NADP(+)-dependent isocitrate dehydrogenase.Planta
   
 
  NADPH kit

Product Code

Unit

Price

Availability

15262

400 assay