AAT Bioquest

Amplite® Colorimetric Ammonia Quantitation Kit *Blue Color*

Ammonia dose response on a 96-well white clear bottom plate using a Spectrum Max microplate reader (Molecular Devices) measured with Amplite® Colorimetric Ammonia/Ammonium Quantitation Kit.
Ammonia dose response on a 96-well white clear bottom plate using a Spectrum Max microplate reader (Molecular Devices) measured with Amplite® Colorimetric Ammonia/Ammonium Quantitation Kit.
Ammonia dose response on a 96-well white clear bottom plate using a Spectrum Max microplate reader (Molecular Devices) measured with Amplite® Colorimetric Ammonia/Ammonium Quantitation Kit.
Ordering information
Catalog Number
Unit Size
Add to cart
Additional ordering information
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
ShippingStandard overnight for United States, inquire for international
Request quotation
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22


Ammonia is an important source of nitrogen for living systems. It is synthesized through amino acid metabolism and is toxic when present at high concentrations. It is produced in liver and converted to urea through the urea cycle. Elevated levels of ammonia in the blood (hyperammonemia) have been found in liver dysfunction (cirrhosis), while hypoammonemia has been associated with defects in the urea cycle enzymes (e.g. ornithine transcarbamylase). The determination of ammonia is very useful test in clinical laboratory to monitor health status. Our Amplite® Colorimetric Ammonia Assay Kit provides a simple and sensitive colorimetric method for the quantitation of ammonia concentration in foods and biological samples such as serum, plasma and urine, etc. The assay is based on an enzyme-coupled reaction of ammonia in the assay buffer, and finally produces a blue colored product. The intensity of color produced is proportional to the concentration of ammonia in the sample, which can be measured colorimetrically at 660-670 nm. This Amplite® Colorimetric Ammonia Assay Kit provides a simple assay to detect ammonia. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step.


Absorbance microplate reader

Absorbance660 - 670 nm
Recommended plateClear bottom


Example protocol


Protocol summary

  1. Prepare Ammonium Chloride standards or test samples (50 µL)
  2. Add Assay Buffer I (50 µL)
  3. Incubate at RT or 37°C for 5 min
  4. Add Assay Buffer II (50 µL)
  5. Incubate at RT for 30 - 60 min
  6. Monitor Absorbance increase at 660 - 670 nm

Important notes
Thaw all the kit components to room temperature before starting the experiment.


Ammonium Chloride standard

For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/10059

Add 1 µL of 1.0 M Ammonium Chloride Standard (Component C) into 999 µL of DPBS to generate 1000 µM Ammonium Chloride standard solution (AS7). Take 1000 µM Ammonium Chloride standard solution (AS7) and perform 1:3 serial dilutions to get serially diluted Ammonium Chloride standards (AS6 - AS1) with DPBS.


Table 1. Layout of Ammonium Chloride standards and test samples in a clear bottom 96-well microplate. AS= Ammonium Chloride Standards (AS1 - AS7, 1 to 1000 µM), BL=Blank Control, TS=Test Samples. 


Table 2. Reagent composition for each well.

AS1 - AS750 µLSerial Dilutions (1 to 1000 µM)
TS50 µLtest sample
  1. Prepare Ammonium Chloride standards (AS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 25 µL of reagent per well instead of 50 µL.

  2. Add 50 µL of Assay Buffer I (Component A) to each well of Ammonium Chloride standard, blank control, and test samples to make the total assay volume of 100 µL/well. For a 384-well plate, add 25 µL of Assay Buffer I into each well instead, for a total volume of 50 µL/well.

  3. Incubate the reaction at room temperature or 37°C for 5 minutes.

  4. Add 50 µL of Assay Buffer II (Component B) to each well to make the total assay volume of 150 µL/well. For a 384-well plate, add 25 µL of Assay Buffer II into each well instead, for a total volume of 75 µL/well.

  5. Incubate the reaction at room temperature for 30 - 60 minutes.

  6. Monitor the absorbance increase with an absorbance microplate reader at 660 - 670 nm. Note: The color turns to yellow after Assay Buffer II (Component B) is added, and the wells with Ammonium Chloride standard or samples will show bluish green color after incubation. The intensity of the color will reach the maximum in 30 - 60 minutes.



View all 4 citations: Citation Explorer
Hepatic ketone body regulation of renal gluconeogenesis
Authors: Hatano, Ryo and Lee, Eunyoung and Sato, Hiromi and Kiuchi, Masahiro and Hirahara, Kiyoshi and Nakagawa, Yoshimi and Shimano, Hitoshi and Nakayama, Toshinori and Tanaka, Tomoaki and Miki, Takashi
Journal: Molecular Metabolism (2024): 101934
A high-protein diet-responsive gut hormone regulates behavioural and metabolic optimization in Drosophila melanogaster
Authors: Niwa, Ryusuke and Yoshinari, Yuto and Nishimura, Takashi and Yoshii, Taishi and Kondo, Shu and Tanimoto, Hiromu and Kobayashi, Tomoe and Matsuyama, Makoto
Journal: (2024)
High-Aspect-Ratio SU-8-Based Optofluidic Device for Ammonia Detection in Cell Culture Media
Authors: Dervisevic, Esma and Voelcker, Nicolas H and Risbridger, Gail and Tuck, Kellie L and Cadarso, Victor J
Journal: ACS sensors (2020)
The use of controls for consistent and accurate measurements of electrocatalytic ammonia synthesis from dinitrogen
Authors: Greenlee, Lauren F and Renner, Julie N and Foster, Shelby L
Journal: (2018)


View all 173 references: Citation Explorer
Systemic inflammation and ammonia in hepatic encephalopathy
Authors: Tranah TH, Vijay GK, Ryan JM, Shawcross DL.
Journal: Metab Brain Dis (2013): 1
High pleural ammonia negatively interferes with the measurement of adenosine deaminase activity
Authors: Loh TP, Tan KM, Chew S, Chan DS.
Journal: BMJ Case Rep (2013)
Role of branched chain amino acids in cerebral ammonia homeostasis related to hepatic encephalopathy
Authors: Bak LK, Waagepetersen HS, Sorensen M, Ott P, Vilstrup H, Keiding S, Schousboe A.
Journal: Metab Brain Dis. (2013)
Multiple functions of the crustacean gill: osmotic/ionic regulation, acid-base balance, ammonia excretion, and bioaccumulation of toxic metals
Authors: Henry RP, Lucu C, Onken H, Weihrauch D.
Journal: Front Physiol (2012): 431
Catalytic mechanisms and biocatalytic applications of aspartate and methylaspartate ammonia lyases
Authors: de Villiers M, Puthan Veetil V, Raj H, de Villiers J, Poelarends GJ.
Journal: ACS Chem Biol (2012): 1618
Mechanistic aspects and elementary steps of N-H bond activation of ammonia and C-N coupling induced by gas-phase ions: a combined experimental/computational exercise
Authors: Kretschmer R, Schlangen M, Schwarz H.
Journal: Chemistry (2012): 40
More actors in ammonia absorption by the thick ascending limb
Authors: Houillier P, Bourgeois S.
Journal: Am J Physiol Renal Physiol (2012): F293
The role of glutamate dehydrogenase in mammalian ammonia metabolism
Authors: Spanaki C, Plaitakis A.
Journal: Neurotox Res (2012): 117
Seven things fish know about ammonia and we don't
Authors: Wright PA, Wood CM.
Journal: Respir Physiol Neurobiol (2012): 231
Ammonia-lowering strategies for the treatment of hepatic encephalopathy
Authors: Rose CF., undefined
Journal: Clin Pharmacol Ther (2012): 321