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Amplite® Colorimetric Alanine Aminotransferase Assay Kit *Blue Color*

ALT dose response was measured with Amplite Colorimetric Alanine Aminotransferase Assay Kit in a 96-well white, clear-bottom plate using a SpectraMax microplate reader (Molecular Devices).
ALT dose response was measured with Amplite Colorimetric Alanine Aminotransferase Assay Kit in a 96-well white, clear-bottom plate using a SpectraMax microplate reader (Molecular Devices).
<strong>Analysis of serum transaminases following injection with viruses.</strong><br>(A) ALT concentration following injection with AdEasy, AdEasy-HMGA-6 or PBS as vehicle control into the liver. (B) ALT concentration following injection with AdEasy, AdEasy-HMGA-6 or PBS as vehicle control into the pancreas. (C) AST concentration following injection with AdEasy, AdEasy-HMGA-6 or PBS as vehicle control into the liver. (D) AST concentration following injection with AdEasy, AdEasy-HMGA-6 or PBS as vehicle control into the pancreas. Viruses were injected with a dose of 1.0X108 virus particles / kg of body weight. 20μL of PBS was injected as a vehicle control. The sham control group did not undergo any surgical procedure. Serum samples were collected 6h, 3, 7 or 30 days post-injection of viruses or PBS. All data represent the means ± SD of three mice. Source: <strong>A mouse model study of toxicity and biodistribution of a replication defective adenovirus serotype 5 virus with its genome engineered to contain a decoy hyper binding site to sequester and suppress oncogenic HMGA1 as a new cancer treatment therapy, </strong>by Faizule Hassan et al., <em>PLOS</em>, Feb. 2018
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Catalog Number13803
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InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22


Alanine aminotransferase (ALT), also called serum glutamate pyruvic transaminase (GPT), is a member of transferase family. It catalyzes the reversible transfer of an alpha-amino group between alanine and glutamate, and is an important enzyme in amino acid metabolism. ALT is found mainly in liver and small amount in heart, muscle, and kidneys. In healthy subjects, serum ALT levels are low. However, when cells are damaged, such as acute and chronic hepatitis, obstructive jaundice, carcinoma of liver, myocardial infarction, ALT may leak into the blood stream and the ALT levels are significantly elevated. Therefore, determination of serum ALT level has great clinical and diagnostic significance. Amplite® Colorimetric Alanine Aminotransferase Assay Kit provides a quick and sensitive method for the measurement of ALT in various biological samples. ALT catalyzes the reaction of alanine and α-ketoglutarate to pyruvate and glutamate. The product glutamate is measured by the generation of a blue color product through an enzyme coupled reaction cycle. The signal can be read by an absorbance microplate reader at an absorbance ratio of A570 nm to A610 nm. With the Amplite® Colorimetric Alanine Aminotransferase Assay Kit as little as 10 mU/mL ALT was detected in a 100 µL reaction volume. The assay is robust, and can be readily adapted for a wide variety of applications.


Absorbance microplate reader

Absorbance570/610 nm
Recommended plateClear bottom


Component A: ALT Enzyme Mixture1 bottle (lyophilized powder)
Component B: ALT Assay Bufferbottle (10 mL)
Component C: NAD1 vial
Component D: ALT Positive Control1 vial (10 U)

Example protocol


Protocol Summary
  1. Prepare ALT working solution (50 µL)
  2. Add ALT standards or test samples (50 µL)
  3. Incubate at 37 °C for 60 mins to 120 mins
  4. Monitor absorbance increase at the absorbance ratio of A570nm/A610nm 
Important      Thaw one bottle Component A and B at room temperature before starting the experiment.


Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

1. ALT standard solution (100 U/mL)
Add 100 µL DPBS into the vial of ALT Positive Control (Component D) to make 100 U/mL ALT standard solution.

2. NAD solution (100X)
Add 100 µL of ddH2O into the vial of NAD (Component C) to have 100X NAD solution.


For convenience, use the Serial Dilution Planner:

ALT standard
Add 10 µL of 100 U/mL ALT standard solution into 990 µL DPBS buffer with 0.1% BSA to generate 1 U/mL ALT standard solution (ALT7). Take 1 U/mL ALT standard solution and perform 1:2 serial dilutions in DPBS buffer with 0.1% BSA to get serial dilutions of ALT standard (ALT6 - ALT1).


  1. Add 10 mL of ALT Assay Buffer (Component B) into the bottle of ALT Enzyme Mixture (Component A) and mix well.
  2. Add the whole vial of 100X NAD solution into the ALT Enzyme Mixture solution to make ALT working solution.
    Note      This ALT working solution is enough for two 96-well plates. It is unstable at room temperature, and should be used promptly within 2 hours and avoid exposure to light. Alternatively, one can make a 50X of ALT Enzyme Mixture stock solution by adding 200 μL of H2O into the bottle of Component A, and then prepare the ALT working solution by mix the stock solution with assay buffer (Component B) and 100X NAD solution proportionally.  


Table 1. Layout of ALT standards and test samples in a clear, white, or black with clear bottom 96-well microplate. ALT= ALT Standards (ALT1 - ALT7, 15.6 to 1000 mU/mL), BL=Blank Control, TS=Test Samples
Table 2. Reagent composition for each well.
ALT1 - ALT750 µLSerial Dilutions (15.6 to 1000 mU/mL)
BL50 µLDPBS with 0.1% BSA
TS50 µLtest sample
  1. Prepare ALT standards (ALT), 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.
    Note      Dilute the test samples to the concentration range in DPBS buffer with 0.1% BSA if needed.
  2. Add 50 µL of ALT working solution to each well of ALT standard, blank control, and test samples to make the total ALT assay volume of 100 µL/well. For a 384-well plate, add 25 µL of ALT working solution into each well instead, for a total volume of 50 µL/well.
  3. Incubate the reaction at 37°C for 60 min to 120 minutes, protected from light. 
    Note      The background of Blank Control increases with time.
  4. Monitor the absorbance increase with an absorbance plate reader at the absorbance ratio of A570nm/A610nm


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Pharmacologic IRE1/XBP1s activation promotes systemic adaptive remodeling in obesity
Authors: Madhavan, Aparajita and Kok, Bernard P and Rius, Bibiana and Grandjean, Julia and Alabi, Adekunle and Albert, Verena and Sukiasyan, Ara and Powers, Evan T and Galmozzi, Andrea and Saez, Enrique and others,
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Reparative and toxicity-reducing effects of liposome-encapsulated saikosaponin in mice with liver fibrosis
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Journal: Bioscience Reports (2020)
Spermidine confers liver protection by enhancing NRF 2 signaling through a MAP 1S-mediated non-canonical mechanism
Authors: Liu, Pengfei and de la Vega, Montserrat Rojo and Dodson, Matthew and Yue, Fei and Shi, Boyun and Fang, Deyu and Chapman, Eli and Liu, Leyuan and Zhang, Donna D
Journal: Hepatology (2019)
Safety of nanoparticles based on albumin-polymer conjugates as a carrier of nucleotides for pancreatic cancer therapy
Authors: Taguchi, Kazuaki and Lu, Hongxu and Jiang, Yanyan and Hung, Tzong-Tyng and Stenzel, Martina Heide
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