生物测定系统/enzyflou&trade；NAD/NADH检测试剂盒/100次检测/EFND-100
商品编号：
EFND-100
品牌：
bioassaysys
市场价：
¥7780.00
美元价：
3890.00
产品分类：
RNA干扰
公司分类：
RNA_interference
联系Q Q：
3392242852
电话号码：
4000-520-616
电子邮箱：
info@ebiomall.com
商品介绍
EnzyFluo™ NAD/NADH Assay Kit
- Product Overview
- Product FAQ
- Product Citations
- Assay Service
ProtocolSDS
Application
- For sensitive determination of NAD and NADH and evaluation of drug effects on NAD/NADH metabolism. Direct NAD/NADH Assays in 96-Well PlateNEW!!!
Key Features
- Sensitive and accurate. Detection limit of 0.02 μM and linearity up to 1 μM NAD+/NADH in 96-well plate assay.
- Convenient. The procedure involves adding a single working reagent, and reading the fluorescence at time zero and 10 min.
- High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.
Method
- F530/585nm
Samples
- Cell, tissue extracts etc Direct NAD/NADH Assays in 96-Well PlateNEW!!!
Species
- All
Size
- 100 tests
Detection Limit
- 0.02 μM
Shelf Life
- 6 months
More Details
- 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. BioAssay Systems EnzyFluo™ 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/em = 530/585 nm, is proportional to the NAD+/NADH concentration in the sample. This assay is highly specific for NAD+/NADH with minimal interference (<1%) by="">+/NADPH and is a convenient method to measure NAD, NADH and their ratio. Direct NAD/NADH Assays in 96-Well PlateNEW!!!
Can I determine NAD/NADH directly in 96-well plates?
Yes, NAD/NADH can be determined directly in 96-well plate. Please see our optimized protocolDirect NAD/NADH Assays in 96-Well PlateWhen tissue is used, should it be freshly obtained? Or is -80°C storage ok?
If direct sample processing is not possible, we recommend snap freezing tissue samples in liquid nitrogen and keeping them either in liquid nitrogen or at -80°C until further processing.I have conducted protocols that involve trypsinizing cells, washing with PBS, pelleting, and then applying extraction buffer. My concern with this method is that PBS is known to cause mitochondrial fragmentation very soon after application to cells. Obviously, the functionality of the mitochondria is very important for NADH/NAD levels. I developed an alternative by keeping my cells adhered to the plate, washing once with pbs, applying heated extraction buffer to the cells directly and then scraping cells off. Do you have any thoughts or experience that would indicate whether one method is better than the other?
The second method is better because the first method may cause mitochondrial fragmentation and thus may alter intracellular NAD/NADH concentrations. The second method involves less pretreatment. Adding the heated extraction buffer terminates the metabolic reactions in the cells and is thus much less likely to alter the intracellular NAD/NADH concentrations.I have also done many experiments with different numbers of cells. What I notice is that there seems to be a sigmoidal relationship between the number of cells and the NAD and NADH concentrations. I am using C2C12 and H9C2 cells. Do you have any experience with these cells at different seeding densities? Also, do you consistently see a relationship between number of cells NAD(H) concentrations? Is this relationship linear, exponential, sigmoidal?
We have not done these experiments. Your results are very interesting, and not unexpected. In general, cells are healthy at certain densities. When cells lack contacts for a healthy environment at low density, they are likely to produce less NAD/NADH, and if too dense, the cells become less active and thus would reach a plateau in NAD/NADH concentrations.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.Liemburg-Apers, Dania C., et al (2016). Acute stimulation of glucose influx upon mitoenergetic dysfunction requires LKB1, AMPK, Sirt2 and mTOR-RAPTOR. J Cell Sci 129.23: 4411-4423. Assay: NAD/NADH in mouse cells. Kim, Ha-Neui, et al. (2015) Sirtuin1 suppresses osteoclastogenesis by deacetylating FoxOs. Molecular Endocrinology 29.10: 1498-1509. Assay: NAD/NADH in mice.To find more recent publications, pleaseclick here.
If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.Simply send us your samples:- Fast turnaround - Quality data - Low costPlease email or call 1-510-782-9988 x 2 to request assay service.
品牌介绍
BioAssay Systems位于加州北部，是一家专注于提供高质量试剂盒的生物技术公司，试剂盒种类广泛、使用简便、操作简单、快速且性能优越，用户不需花额外的时间来作调适测试。BioAssay Systems产品具有快速、简单、高通量（可用于HTS药物筛选）、灵敏(特别适合小鼠样品) 、准确性高和低干扰等主要特征，且采用发光、生物发光、化学发光、荧光等非放射性的检测技术，安全无害可靠。
