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MAK310

Sigma-Aldrich

Ammonia Assay Kit

Ammonia Assay Kit

sufficient for 200 fluorometric tests

Synonym(s):

Ammonia Quantitative Test Kit

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.

detection method

fluorometric

relevant disease(s)

genitourinary disorders; gastrointestinal diseases

storage temp.

−20°C

General description

Ammonia (NH3) or the ammonium ion (NH4 +) is found in the atmosphere, rainwater, soil, seawater, and volcanic areas. It is widely used as fertilizer, being an important source of nitrogen for living systems. It also plays a role in animal physiology. Ammonia is produced by amino acid metabolism and is converted to urea in the human liver through the urea cycle. Ammonia acts as a precursor for various other nitrogen compounds. It is a colorless gas with pungent odor and can be easily liquefied. It is also used as a feed supplement in livestock industry.[1]

Features and Benefits

Compatible with high-throughput handling systems.

Suitability

Suitable for the quantitative determination of ammonia/ammonium concentration in biological samples, such as urine, and environmental samples

Principle

The Ammonia Assay Kit provides a simple and high throughput adaptable assay for quantitative determination of ammonia/ammonium ion concentration in biological samples, such as urine, and environmental samples. This assay is based on the o-phthalaldehyde method in which the reagent reacts with ammonia/ammonium ion producing a fluorometric result ( ex = 360/ em = 450 nm), proportional to the ammonia concentration in the sample. The kit has a linear detection range of 0.012–1 mM ammonia in a 96 well format.

Signal Word

Danger

Hazard Statements

H302,H314,H317,H410

Hazard Classifications

Acute Tox. 4 Oral - Aquatic Acute 1 - Aquatic Chronic 1 - Eye Dam. 1 - Skin Corr. 1B - Skin Sens. 1

Storage Class Code

8A - Combustible corrosive hazardous materials

Flash Point(F)

188.6 °F

Flash Point(C)

87 °C

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Certificates of Analysis (COA)

Lot/Batch Number
310CD09A28
310CD08A02
310CD04A19
310CC09A20
TEST2_COFA_TEXT

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Daphné Coache et al.
PloS one, 17(6), e0270206-e0270206 (2022-06-25)
This study assessed the stability of six extemporaneously compounded hydroxyurea oral liquids stored at room temperature. Hydroxyurea oral liquids (100 mg/mL) were prepared using three different mixing methods (mortar, mixer or QuartetRx) from either bulk powder, capsule content, or whole
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Excitotoxicity is a cellular phenomenon that comprises the consequences of toxic actions of excitatory neurotransmitters, such as glutamate. This process is usually related to overproduction of reactive oxygen species (ROS) and ammonia (NH4+ ) toxicity. Platinum nanoparticle (Pt-NP)-based microreactors able
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1 Review This action will navigate to reviews.

Questions

1–7 of 7 Questions  

  1. What is the composition of reagent A and reagent B supplied with the MAK310 assay kit?

    1 answer

    1. The exact percentages as exact reagent composition are proprietary. The composition can be provided in ranges as follows; Reagent A: Contains 1-15% o-phthalaldehyde, CAS #: 643-79-8, and 10-99% dimethyl sulfoxide, CAS#: 67-68-5. Reagent B: Contains 0.1-5% β-Mercaptoethanol, CAS#: 60-24-2, and 10-99% dimethyl sulfoxide, CAS#: 67-68-5.

      Helpful?

  2. Can i use this assay kit to determine the ammonia amount of culture media sample? (RPMI-1640)

    1 answer

    1. This kit has not been tested for use with cell culture samples or supernatants, including those containing RPMI-1640. Amine-containing buffers such as Tris buffers should be avoided. In addition, high protein concentrations may interfere with the reaction. The end user would have to determine suitability. Product MAK538, Ammonia Assay Kit, would be a more suitable product for use in cell culture applications. Please see the link below to review this product option:
      https://www.sigmaaldrich.com/product/sigma/mak538

      Helpful?

  3. What is the recommended protocol for the storage and freeze/thaw of the ammonia assay kit (MAK310)? Is it permissible to aliquot portions in advance, or is the kit stable for a certain period of time at 4°C? Furthermore, what is the optimal method for thawing the kit (on ice)? Additionally, what is the shelf life of the reagents once thawed? Should the thawed "working" reagents be stored at 4°C or room temperature until they are used (within the timeframe of the above question)?

    1 answer

    1. It is advisable to aliquot portions of the reagents in advance to minimize the number of freeze-thaw cycles. Thawing the components at room temperature is acceptable. But, there is no specific data available regarding the stability of the reagents once they are thawed. It is recommended to prepare smaller aliquots of the reagents, enough for the number of assays the customer intends to run, to avoid having an excessive amount of reagent after an experiment. While it is acceptable to refreeze the leftover aliquots for future use, it is advisable to minimize multiple freeze-thaw cycles. The reagents may be temporarily stored at 4°C if necessary during an experiment. Additionally, it is recommended to create a standard curve according to the protocol, and if the curve is linear, then the kit should be functioning properly.

      Helpful?

  4. Can this kit chemistry be used directly with serum samples, or will the protein content interfere with the assay?

    1 answer

    1. The kit chemistry cannot be used directly with serum samples. Deproteinization is necessary before running the assay, and this can be achieved by passing the sample through 10 kDa cutoff filters. Alternatively, proteins can be chemically precipitated using TCA, although this method has not been tested with this kit. If using acid for protein separation, it is important to neutralize the sample with a base before running the assay.

      Helpful?

  5. To calculate the molarity of my samples, do I need to multiply it by the dilution factor of the final reaction volume? According to technical bulletin, standards are diluted 1/10 at final molarity of .1 mM at the highest point of the curve.

    1 answer

    1. If 10 uL of undiluted sample is used before adding the 90 uL of working reagent, then no dilution factor is needed. The dilution factor is only needed if the sample has been diluted before the pipetting for 10 uL into the sample well. For example, if the kit is used and the ammonia concentration is 1 mM or greater, it is necessary to dilute the sample in water and repeat the assay. Multiply the results by the dilution factor. If the sample is diluted 10-fold, then a dilution factor of '10' would need to be applied to the results.

      Helpful?

  6. Is this a stand alone test or does it require external instrumentation?

    1 answer

    1. A 96 well plate is not included in this kit. A plate reader is also required. Please see the 'Reagents and Equipment Required but Not Provided with this kit' on the Product Information Sheet:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/196/522/mak310bul.pdf

      Helpful?

  7. Is this kit suitable for seawater samples?

    1 answer

    1. Yes, the Ammonia Assay Kit provides a simple and high throughput adaptable assay for quantitative determination of ammonia/ammonium ion concentration in biological samples, such as urine, and environmental samples.

      Helpful?

Reviews

Active Filters

    • Review 1
    • Votes 0
    2 out of 5 stars.

    Limited uses

    I would not recommend using this kit for any kind of sample that contains free amino acids as o-phthalaldehyde will readily bind and contribute to fluorescence. I tested for a few different types of cell culture media (including protein depleted samples) and also buffer samples containing only glutamate and alanine and all gave dose dependent levels of interference, making the kit very non-specific. However, for ammonia only samples in simple and appropriate buffers, it worked fine.

    Helpful?

    1. Response from MilliporeSigma:

      This assay is based on the o-phthalaldehyde method in which the reagent reacts with ammonia/ammonium ion producing a fluorometric result ( ex = 360/ em = 450nm), proportional to the ammonia concentration in the sample. It will not differentiate between ammonia, free amino acids, or other components that react with o-phthalaldehyde. The end user would need to determine suitability with their samples.

      We encourage customers who experience problems with our products to contact us for additional technical support. Please visit https://www.sigmaaldrich.com/US/en/support/customer-support to submit a Product Technical Inquiry.

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