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Ammonia Colorimetric Assay Kit II

Sensitive Assay, HTS
Catalog #: K470
$415.00

Product Details

Cat # +Size K470-100
Size 100 assays
Detection Method Absorbance (670 nm)
Species Reactivity Mammalian
Applications The kit can detect less than 1 nmol (~10 μM) ammonia using either a 96 well or 384 well microwell plate.
Features & Benefits • Simple procedure; takes ~ 3 hours
• Fast and convenient
• The assay is sensitive and stable
• The TUNEL-based assay kit provides complete components including positive and negative control cells for convenient detection of DNA fragmentation in cultured cells and tissue sections.
Kit Components • Ammonia Reagent 1
• Ammonia Reagent 2
• Ammonium Chloride Standard (100 mM)
Storage Conditions 4°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Ammonia is an important source of nitrogen for living systems. Nitrogen is required for the synthesis of amino acids, which are the building blocks of protein. Ammonia is a metabolic product which is created through amino acid deamination. It plays an important role in both normal and abnormal animal physiology such as acid/base balance. BioVision provides a rapid, simple, sensitive, and reliable assay suitable for research and high throughput assay of Ammonia. In this non-enzymatic assay, ammonia is used to form indophenol, a highly colored product easily quantifiable by colorimetry (OD670 nm) using a plate reader. The kit can detect less than 1 nmol (~10 μM) ammonia using either a 96 well or 384 well microwell plate.


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What substances can interfere with the assay?
Reducing agents beta-mercaptoethanol and DTT can interfere. SDS is not recommended since it denatures the enzymes in the reaction. Glycerol adds density to the solution and hence should be avoided.
What is the difference between K370-100 and K470-100?
Typically for biological samples, K370 is the appropriate one. Pyruvate can create background in this assay. The kit can detect 1 nmol (~20 µM) of ammonia. -K470 forms Indophenol which is indigo/blue in color and is quantified at 670nm. The kit can detect less than 1 nmol (~10 µM) ammonia. Amines, amino acids and hemoglobin interfere with this assay. Typically environmental samples like sea water, soil extracts, etc are suitable for this kit.
Can the kit be used to quantify ammonium of mammalian cell culture supernatant, since the mammalian cell cultures have aminoacids that can interfer with this assay?
Your media can be deproteinized by passing through a 10kDa filter (Cat#1997-25, http://www.biovision.com/10k-spin-column-5277.html) so that all proteins are removed from the medium. The amino acids can generate background readings which can be subtracted using parallel controls. Fig 5 in this reference can help clarify that the reagents used for the BioVision Ammonia assay were selected to minimize interference: E.D. Rhine, G.K. Pratt, R.L. Mulvaney and E.J. Pratt (1998) Soil Sci. Soc. AM. J. 62: 473-480.
The OD values are very high, the standard curve is nonlinear. Why?
High OD is observed when ammonia gets absorbed into the solution/s leading to high values irrespective of the increasing standard concentrations. It is crucial to protect reagents from getting exposure to air. This is a commonly seen issue and has to be avoided with utmost care.
Why can't I creat the same standard curve in tissue culture media?
This is expected because the standard, Ammonium Chloride cannot properly ionize and react because of the buffering property of the medium.
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."
What is the shelf life of this kit?
 BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
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.