NADH and NADPH Assay Kit (Colorimetric) (ab186031) provides a convenient method for detecting NADH and NADPH.
- Colorimetric readout - 460 nm - works with standard microplate readers
- Rapid, no-wash protocol
- Detect as little as 3 µM NADPH in a 100 µL assay volume
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- International journal of molecular sciences 24:2023Opioids Alleviate Oxidative Stress via the Nrf2/HO-1 Pathway in LPS-Stimulated Microglia.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAkash Shivling Mali,Ondrej Honc,Lucie Hejnova,Jiri NovotnyPubMed 37446267
- Nutrients 14:2022Resveratrol in the Hypothalamic Paraventricular Nucleus Attenuates Hypertension by Regulation of ROS and Neurotransmitters.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJie Qi,Li-Yan Fu,Kai-Li Liu,Rui-Juan Li,Jin-An Qiao,Xiao-Jing Yu,Jia-Yue Yu,Ying Li,Zhi-Peng Feng,Qiu-Yue Yi,Hong Jia,Hong-Li Gao,Hong Tan,Yu-Ming KangPubMed 36235829
- Nature communications 13:43272022Targeting de novo lipogenesis and the Lands cycle induces ferroptosis in KRAS-mutant lung cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesCaterina Bartolacci,Cristina Andreani,Gonçalo Vale,Stefano Berto,Margherita Melegari,Anna Colleen Crouch,Dodge L Baluya,George Kemble,Kurt Hodges,Jacqueline Starrett,Katerina Politi,Sandra L Starnes,Daniele Lorenzini,Maria Gabriela Raso,Luisa M Solis Soto,Carmen Behrens,Humam Kadara,Boning Gao,Ignacio I Wistuba,John D Minna,Jeffrey G McDonald,Pier Paolo ScaglioniPubMed 35882862
- Antioxidants (Basel, Switzerland) 11:2022Na/K-ATPase Alpha 2 Isoform Elicits Rac1-Dependent Oxidative Stress and TLR4-Induced Inflammation in the Hypothalamic Paraventricular Nucleus in High Salt-Induced Hypertension.Applications:Unspecified applicationReactive species:Unspecified reactive speciesQing Su,Xiao-Jing Yu,Xiao-Min Wang,Bo Peng,Juan Bai,Hong-Bao Li,Ying Li,Wen-Jie Xia,Li-Yan Fu,Kai-Li Liu,Jin-Jun Liu,Yu-Ming KangPubMed 35204171
- Frontiers in pharmacology 12:7566712021Bilateral Paraventricular Nucleus Upregulation of Extracellular Superoxide Dismutase Decreases Blood Pressure by Regulation of the NLRP3 and Neurotransmitters in Salt-Induced Hypertensive Rats.Applications:Unspecified applicationReactive species:Unspecified reactive speciesQing Su,Xiao-Jing Yu,Xiao-Min Wang,Hong-Bao Li,Ying Li,Juan Bai,Jie Qi,Nianping Zhang,Kai-Li Liu,Yan Zhang,Guo-Qing Zhu,Yu-Ming KangPubMed 34899311
- Biomedicines 8:2020Dibenzoylthiamine Has Powerful Antioxidant and Anti-Inflammatory Properties in Cultured Cells and in Mouse Models of Stress and Neurodegeneration.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMargaux Sambon,Anna Gorlova,Alice Demelenne,Judit Alhama-Riba,Bernard Coumans,Bernard Lakaye,Pierre Wins,Marianne Fillet,Daniel C Anthony,Tatyana Strekalova,Lucien BettendorffPubMed 32962139
- Molecular medicine reports 22:964-9742020Protective effect of microRNA‑340‑5p against oxygen‑glucose deprivation/reperfusion in PC12 cells through targeting neuronal differentiation 4.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJuan Wang et. al.PubMed 32468054
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue, Cell culture supernatant, Cell Lysate
- Assay type
- Quantitative
- Range
- 3.13 - 100 µM
- Assay time
- 2h
- Sensitivity
- > 3 µM
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Target data
Additional Targets
Disodium nicotinamide adenine dinucleotide
What's included?
Recommended products
NADH and NADPH Assay Kit (Colorimetric) (ab186031) provides a convenient method for detecting NADH and NADPH.
- Colorimetric readout - 460 nm - works with standard microplate readers
- Rapid, no-wash protocol
- Detect as little as 3 µM NADPH in a 100 µL assay volume
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue, Cell culture supernatant, Cell Lysate
- Assay type
- Quantitative
- Range
- 3.13 - 100 µM
- Assay time
- 2h
- Assay Platform
- Microplate reader
- Sensitivity
- > 3 µM
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
NADH and NADPH Assay Kit (ab186031) provides a convenient method for detecting NADH and NADPH.
How the NADH / NADPH assay works
Nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) are two important cofactors found in cells. NADH is the reduced form of NAD+. NAD forms NADP with the addition of a phosphate group to the 2' position of the adenyl nucleotide through an ester linkage. The traditional NAD/NADH and NADP/NADPH assays are based on monitoring the changes in NADH or NADPH absorption at 340 nm. The short UV wavelength of NAD/NADH and NADP/NADPH assays makes the traditional methods suffer low sensitivity and high interference.
In this NADH and NADPH assay method, the NADH/NADPH probe is a chromogenic sensor that has its maximum absorbance at 460 nm upon NADH/NADPH reduction. The absorbance maximum increases to ~ 635 nm if the enhancer is added to the assay system. The absorption of the NADH/NADPH probe is directly proportional to the concentration of NADH/NADPH in the solution. Note that this assay kit will measure both NADPH and NADH together, and cannot differentiate them.
The Colorimetric NADPH Assay Kit provides a sensitive assay to detect as little as 3 µM NADPH in a 100 µL assay volume.
NADH and NADPH assay protocol summary
- prepare NADPH Reaction Mixture
- add NADPH standards or test samples
- incubate at room temperature for 15-120 minutes
- monitor Absorbance at 460 nm
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
NADH and NADPH along with their corresponding oxidized forms NAD+ and NADP+ function as critical coenzymes in cellular metabolism. These molecules serve as electron carriers in various biochemical reactions. NADH is primarily involved in energy production through the electron transport chain while NADPH functions mainly in reductive biosynthesis and antioxidant defense. Nadph has a molar mass of approximately 745.41 g/mol. Both are expressed in virtually all living cells due to their role in fundamental cellular processes.
- Biological function summary
These coenzymes play distinct and integral roles in cellular metabolism. NADH primarily supports ATP generation by donating electrons to the mitochondria which culminate in the production of adenosine triphosphate (ATP). Conversely NADPH is essential in biosynthetic processes such as fatty acid and nucleic acid synthesis and it provides reducing power to counteract oxidative stress. These functions illustrate the importance of NADH and NADPH in maintaining cellular homeostasis and supporting anabolic reactions.
- Pathways
NADH and NADPH are key participants in several vital pathways. In cellular respiration NADH is a central actor in glycolysis and the citric acid cycle where it facilitates the transfer of electrons to the electron transport chain enhancing oxidative phosphorylation. On the other hand NADPH serves a predominant role in the pentose phosphate pathway where it delivers reducing power for fatty acid synthesis and the reduction of glutathione. These pathways highlight their interplay with enzymes such as glucose-6-phosphate dehydrogenase and complex I of the mitochondria.
- Associated diseases and disorders
Disruptions in NADH and NADPH homeostasis can lead to various conditions including cancer and neurodegenerative diseases. Cancer cells often exhibit altered glucose metabolism known as the Warburg effect where NADPH supports increased biosynthesis and redox balance. Furthermore neurodegenerative diseases like Parkinson's are linked to mitochondrial dysfunction and oxidative stress where NADH and NADPH levels may become dysregulated. The sirtuin family of proteins which rely on NAD+ availability also highlight a connection between these coenzymes and disease pathophysiology.
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2 product images
Functional Studies - NADH and NADPH Assay Kit (Colorimetric) (ab186031)
NADPH standard curve.
NADPH dose response was measured with the Colorimetric NADPH Assay Kit in a 96-well white/clear bottom plate using a microplate reader. As low as 3 μM of NAPDH can be detected with 30min incubation (n=3) with absorbance measurement at 460nm.
The absorbance in blank wells (with the PBS buffer only) is used as a control, and is subtracted from the values for those wells with the NADPH reactions.
Functional Studies - NADH and NADPH Assay Kit (Colorimetric) (ab186031)
NADH dose response curve.
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