Amplite® Colorimetric Ammonia Quantitation Kit *Blue Color*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12171501 |
Overview | ![]() ![]() |
See also: Amplite® Reagents and Assay Kits, Cell Metabolism
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.
Platform
Absorbance microplate reader
Absorbance | 660 - 670 nm |
Recommended plate | Clear bottom |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare Ammonium Chloride standards or test samples (50 µL)
- Add Assay Buffer I (50 µL)
- Incubate at RT or 37°C for 5 min
- Add Assay Buffer II (50 µL)
- Incubate at RT for 30 - 60 min
- Monitor Absorbance increase at 660 - 670 nm
Important notes
Thaw all the kit components to room temperature before starting the experiment.
PREPARATION OF STANDARD SOLUTION
Ammonium Chloride standard
For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/10059
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.
SAMPLE EXPERIMENTAL PROTOCOL
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.
BL | BL | TS | TS |
AS1 | AS1 | ... | ... |
AS2 | AS2 | ... | ... |
AS3 | AS3 | ||
AS4 | AS4 | ||
AS5 | AS5 | ||
AS6 | AS6 | ||
AS7 | AS7 |
Table 2. Reagent composition for each well.
Well | Volume | Reagent |
AS1 - AS7 | 50 µL | Serial Dilutions (1 to 1000 µM) |
BL | 50 µL | DPBS |
TS | 50 µL | test sample |
- 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.
- 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.
- Incubate the reaction at room temperature or 37°C for 5 minutes.
- 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.
- Incubate the reaction at room temperature for 30 - 60 minutes.
- 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.
Images
Citations
View all 2 citations: Citation Explorer
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)
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)
Authors: Greenlee, Lauren F and Renner, Julie N and Foster, Shelby L
Journal: (2018)
References
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
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)
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)
Authors: Bak LK, Waagepetersen HS, Sorensen M, Ott P, Vilstrup H, Keiding S, Schousboe A.
Journal: Metab Brain Dis. (2013)
Ammonia transport by terrestrial and aquatic insects
Authors: Weihrauch D, Donini A, O'Donnell MJ.
Journal: J Insect Physiol (2012): 473
Authors: Weihrauch D, Donini A, O'Donnell MJ.
Journal: J Insect Physiol (2012): 473
Physiology and diversity of ammonia-oxidizing archaea
Authors: Stahl DA, de la Torre JR.
Journal: Annu Rev Microbiol (2012): 83
Authors: Stahl DA, de la Torre JR.
Journal: Annu Rev Microbiol (2012): 83
Effect of Helicobacter pylori eradication on blood ammonia levels in cirrhotic patients: a systematic review
Authors: Qin SY, Jiang HX, Ning HJ, Nie HM, Tao L, Hu BL, Guo XY.
Journal: Hepatogastroenterology (2012): 2576
Authors: Qin SY, Jiang HX, Ning HJ, Nie HM, Tao L, Hu BL, Guo XY.
Journal: Hepatogastroenterology (2012): 2576
Drivers of archaeal ammonia-oxidizing communities in soil
Authors: Zhalnina K, de Quadros PD, Camargo FA, Triplett EW.
Journal: Front Microbiol (2012): 210
Authors: Zhalnina K, de Quadros PD, Camargo FA, Triplett EW.
Journal: Front Microbiol (2012): 210
Is there in vivo evidence for amino acid shuttles carrying ammonia from neurons to astrocytes
Authors: Rothman DL, De Feyter HM, Maciejewski PK, Behar KL.
Journal: Neurochem Res (2012): 2597
Authors: Rothman DL, De Feyter HM, Maciejewski PK, Behar KL.
Journal: Neurochem Res (2012): 2597
A review of ammonia-oxidizing bacteria and archaea in Chinese soils
Authors: Shen JP, Zhang LM, Di HJ, He JZ.
Journal: Front Microbiol (2012): 296
Authors: Shen JP, Zhang LM, Di HJ, He JZ.
Journal: Front Microbiol (2012): 296
Ammonia-lowering strategies for the treatment of hepatic encephalopathy
Authors: Rose CF., undefined
Journal: Clin Pharmacol Ther (2012): 321
Authors: Rose CF., undefined
Journal: Clin Pharmacol Ther (2012): 321
Application notes
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