This kit has not been tested for use on bacterial cultures and may not be suitable. It would be up to the end user to determine suitability. However, the colorimetric version of this kit, MAK468, has been successfully used in Corynebacterium glutamicum, Klebsiella pneumoniae, Lactobacillus plantarum, Edwardsiella tarda, Acinetobacter baumannii, Auxenochlorella protothecoides, E. coli, and other microbial species. Please see the links below to review this product and protocol.
MAK468 product page:
https://www.sigmaaldrich.com/product/sigma/mak468
MAK468 datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/219/480/mak468pis-ms.pdf
MAK460
NAD+/NADH Assay Kit
Sufficient for 100 fluorometric tests
Synonym(s):
NAD/NADH Quantitation Kit
Sign Into View Organizational & Contract Pricing
All Photos(1)
About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.
Recommended Products
Quality Level
application(s)
pharmaceutical
detection method
fluorometric
relevant disease(s)
cancer, Alzheimer′s disease; neurological disorders, Parkinson′s disease; aging/geriatric diseases
storage temp.
−20°C
General description
Pyridine nucleotides play an important role in metabolism and, thus, there is continual interest in monitoring their concentration levels. Quantitative determination of NAD+/NADH has applications in research pertaining to energy transformation and redox state of cells or tissue.
Pyridine nucleotides, including nicotinamide adenine dinucleotide (NAD), are crucial in cellular metabolism, impacting how energy is processed and managed within cells. Monitoring the concentrations of NAD and its reduced form, NADH, is vital for understanding cellular energy dynamics and redox balance. NAD, functioning as a central electron carrier, alternates between NAD+ (oxidized) and NADH (reduced) forms, facilitating numerous metabolic reactions. This transformation is essential not only for redox reactions but also for other metabolic functions, such as ADP-ribosylation, which is crucial for regulating protein functions and DNA repair. Additionally, NAD+ acts as a substrate for sirtuins, which are important for energy metabolism and modulating oxidative stress. .
Application
- Enhanced glycolysis in the myometrium with ectopic endometrium of patients with adenomyosis: a preliminary study.: This study investigates the role of glycolysis in the myometrium of patients with adenomyosis, utilizing the NAD+/NADH Assay Kit to assess metabolic alterations (Huang et al., 2024). 10.1080/09513590.2024.2332411
- Jian-Pi-Yi-Shen formula alleviates renal fibrosis by restoring NAD+ biosynthesis in vivo and in vitro.: This research demonstrates how the Jian-Pi-Yi-Shen formula mitigates renal fibrosis through NAD+ biosynthesis, with the NAD+/NADH Assay Kit playing a crucial role in the biochemical analysis (Gao et al., 2023). 10.18632/aging.205352
- Butyrate inhibits the mitochondrial complex Ι to mediate mitochondria-dependent apoptosis of cervical cancer cells.: The study explores butyrate′s effects on mitochondrial function and apoptosis in cervical cancer cells, employing the NAD+/NADH Assay Kit to measure mitochondrial activity (Zhang et al., 2023). 10.1186/s12906-023-04043-3
- PM2.5 mediates mouse testis Sertoli TM4 cell damage by reducing cellular NAD().: This article investigates the impact of PM2.5 on Sertoli cells in mouse testes, using the NAD+/NADH Assay Kit to track changes in cellular NAD levels (Xu et al., 2023). 10.1080/15376516.2023.2215862
- Synergistic Effect and Mechanism of Plumbagin with Gentamicin Against Carbapenem-Resistant Klebsiella pneumoniae.: The research highlights the synergistic antibacterial effects of plumbagin and gentamicin, with the NAD+/NADH Assay Kit being used to evaluate the metabolic impact on bacterial cells (Chen et al., 2020). 10.2147/IDR.S265753
The NAD+/NADH Assay Kit may be used for:
- Cancer Research
- Neurodegenerative Research
- Age/Geriatric Related Disease Research
Features and Benefits
Broad Detection Range: Detect NAD and NADH levels accurately across a wide range, from 0.02 to 1 μM, using only 50 μL of sample in a 96-well plate setup, ensuring comprehensive analysis with minimal sample requirement
Simplified Process: Experience a streamlined process with the addition of only a single working reagent and a 10 minute room temperature reaction, reducing complexity and saving valuable time and effort.
Compatibility with High-Throughput Systems: Easily incorporate our kit into high-throughput handling systems, ensuring smooth and accurate processing, enhancing efficiency in your laboratory workflow.
Simplified Process: Experience a streamlined process with the addition of only a single working reagent and a 10 minute room temperature reaction, reducing complexity and saving valuable time and effort.
Compatibility with High-Throughput Systems: Easily incorporate our kit into high-throughput handling systems, ensuring smooth and accurate processing, enhancing efficiency in your laboratory workflow.
Suitability
The kit is suitable for the determination of NAD and NADH and the evaluation of drug effects on NAD/NADH metabolism in cells, tissue extracts etc.
Principle
The NAD+/NADH assay kit is based on a lactate dehydrogenase cycling reaction, in which the formed NADH reduces a probe into a highly fluorescent product. The fluorescence intensity of this product, measured at λEx = 530 nm/λEm = 585 nm, is proportional to the NAD+/NADH concentration in the sample. This assay is highly specific for NAD+/NADH with minimal interference (<1%) by NADP+/NADPH and is a convenient method to measure NAD, NADH and their ratio.
Other Notes
For additional information on our range of Biochemicals, please complete this form.
Signal Word
Warning
Hazard Statements
Precautionary Statements
Hazard Classifications
Met. Corr. 1
Storage Class Code
8B - Non-combustible corrosive hazardous materials
Choose from one of the most recent versions:
Certificates of Analysis (COA)
Lot/Batch Number
Don't see the Right Version?
If you require a particular version, you can look up a specific certificate by the Lot or Batch number.
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
-
Can this kit be used for detection of NADH levels in bacteria culture?
1 answer-
Helpful?
-
-
Is there any antibody epitope information available for the products IL-12p40 and IL-12p70?
1 answer-
For IL-12p40, the capture and detection antibodies are mouse monoclonal anti-human IL-12p40. As for IL-12p70, the capture antibody is rat monoclonal anti-human IL-12p70, and the detection antibody is mouse monoclonal anti-human IL-12p70. However, there is no available information on antibody binding domains.
Helpful?
-
-
Can a white plate be used instead of a black plate for this assay?
1 answer-
A black, opaque, flat-bottomed plate is recommended for fluorescence assays, and a black plate with a clear bottom is also acceptable. White plates are not recommended for fluorescence assays as they can increase background and cross-talk. An example of a recommended plate type for fluorescence assays is the product M9685.
Helpful?
-
-
Does this assay measure total NAD+/NADH concentrations? If so, How can the conversion to NAD+ or NADH only be achieved?
1 answer-
The calculation method is the same for NAD+ and NADH. However, there should be one well for the NAD+ test and another well for the NADH test. If the intention is to solely measure NAD+, the sample should be processed according to the NAD+ instructions. This involves homogenizing the sample with NAD+ extraction buffer, then proceeding with heating and addition of the assay buffer followed by the addition of the opposite buffer (in this case, NAD extraction buffer). The order of addition of the extraction buffers determines whether NAD+ or NADH is being measured, as it degrades the other due to the extraction pH.
Helpful?
-
-
Is this kit's sample only for cultured cells? Do you have any data or protocols for serum, urine, saliva, etc.?
1 answer-
This kit has not been tested with serum or urine samples. Unfortuantely, publications regarding this application could not found.
However, there may not be any problems with using plasma, as well as serum, with the kit. It is prudent to test a number of dilutions of urine to determine the best concentration to use in the assay. The following substances interfere with the assay and should be avoided in sample preparation: EDTA (>0.5 mM), ascorbic acid, SDS (>0.2%), sodium azide, NP-40 (>1%) and Tween-20 (>1%).Helpful?
-
Active Filters
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service
