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Product Details
| Cat # +Size | K208-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Detection method- fluorescence microscope (Ex/Em = 360/450) |
| Applications | Screening/studying/characterizing potential NOS inhibitors |
| Features & Benefits | • Simple protocol. Fluorescent detection |
| Kit Components | • NOS Assay Buffer • NOS Dilution Buffer • NOS Substrate • NOS Cofactor 1 • NOS Cofactor 2 (25X) • Nitrate Reductase • NOS (Positive Control) • Enhancer • Nitrite Standard • Probe • NaOH |
| Storage Conditions | -80°C |
| Shipping Conditions | Dry Ice |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Nitric oxide synthases (EC 1.14.13.39) (NOS) is a family of enzymes that catalyze the production of nitric oxide (NO). In presence of NADPH, FAD, FMN, (6R)-5,6,7,8-tetrahydrobiopterin, calmodulin and heme, NOS catalyzes a five-electron oxidation of the guanidino nitrogen of L-arginine with molecular oxygen to generate NO and L-citrulline. Nitric oxide plays an important role in neurotransmission, vascular regulation, immune response and apoptosis. In contrast to its beneficial effects, NO has also been associated with numerous pathological situations such as hypotension accompanying septic shock, essential hypertension, and atherosclerosis. The overproduction of NO has been found to be the fundamental cause underlying neurodegenerative disorders and neuropathic pain. Therefore, developing small molecules for inhibition of NOS is therapeutically desirable. BioVision’s Nitric Oxide Synthase Inhibitor Screening Kit provides a simple and high-throughput adaptable method to screen/study/characterize potential NOS inhibitors. In this assay, nitric oxide generated by NOS undergoes a series of reactions and reacts with the fluorescent probe to generate a stable signal at Ex/Em = 360/450 nm, which is directly proportional to NOS activity. In the presence of a NOS-specific inhibitor, the formation of NO is reduced/abolished resulting in decrease or total loss of the fluorescence.
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Can the kit be used on muscle tissue homogenates specifically?
The kit can be used with most tissue extracts, provided that the tissues are well homogenized and contains >5µU of NOS activity.
There are 3 isoforms of NOS. Which isoform is included in the positive control?
The positive control is an inducible mouse NOS expressed and purified from E.Coli.
Can this kit work with live cells?
Unfortunately, this is an endpoint assay where the cells need to be lysed. However, our kit K207-100 let's you determine intracellular NOS activity.
Can we use frozen samples with this assay?
Although we recommend using fresh samples, but the assay can work with frozen samples as well.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
