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NOVOCIB’s NADH + NADPH Bioluminescent Kit provides an ultrasensitive, one-step assay to detect as low as 2.10-13 mole of NAD(P)H in a 100µL assay volume (2nM). NOVOCIB’s NADH + NADPH Bioluminescent Kit presents the following key-advantages: • reproducibility and reliability (linearity over 3 orders of magnitude) • a high sensitivity (>0.2 pmoles) • fast and easy to use for immediate measurements. |
New!
Number of colorimetric and fluorometric methods have been developed for detection of micromolar and submicromolar concentrations of NADH and NADPH. The NADH + NADPH Bioluminescent Kit provided by NOVOCIB allows the detection of nanomolar concentrations of NADH + NADPH.
With NOVOCIB's NADH + NADPH Bioluminescent Kit, ultrasenstive quantification of NAD(P)H is done with an enzymatic system consisting of highly pure bioluminescence enzymes: luciferase from Photobacterium phosphoreum (lux) and FMN-oxidoreductase from E.coli (Fre) In this coupled reaction NADH and NADPH are first used by bacterial FMN-oxidoreductase to produce FMNH2. Bacterial luciferase then catalyzes the oxidation of FMNH2 to FMN in the presence of O2 and of a long chain aldehyde, with the emission of light (λ = 490 nm) whose amount is directly proportional to the total concentration of NADH and NADPH:
By carefully controlling the purity of its bioluminescence enzymes and the luciferase / reductase ratio (see our Ref. E-Nov 8 & E-Nov10), NOVOCIB has improved the intensity and light kinetics of NAD(P)H-dependent luminescence resulting in a reliable and ultrasensitive assay. NOVOCIB's kit shows a stable signal over time when compare to conventional bacterial luciferase. Measures were done at different concentration of NADH, in 100µL assay, with a Sirius L luminometer (Berthold). NADH + NADPH Bioluminescent Kit is intended for the quantification of a constant concentration of total NADH and NADPH in the range of 2nM to 1µM.
With NOVOCIB's NADH + NADPH Bioluminescent Kit, ultrasenstive quantification of NAD(P)H is done with an enzymatic system consisting of highly pure bioluminescence enzymes: luciferase from Photobacterium phosphoreum (lux) and FMN-oxidoreductase from E.coli (Fre) In this coupled reaction NADH and NADPH are first used by bacterial FMN-oxidoreductase to produce FMNH2. Bacterial luciferase then catalyzes the oxidation of FMNH2 to FMN in the presence of O2 and of a long chain aldehyde, with the emission of light (λ = 490 nm) whose amount is directly proportional to the total concentration of NADH and NADPH:
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References (with links to PubMed) |
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