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Amplite® Fluorimetric Oxaloacetate Assay Kit *Red Fluorescence*

Oxaloacetate dose response was measured with the Amplite® Fluorimetric Oxaloacetate Assay Kit on a solid black 96-well plate using a Gemini microplate reader (Molecular Devices).
Oxaloacetate dose response was measured with the Amplite® Fluorimetric Oxaloacetate Assay Kit on a solid black 96-well plate using a Gemini microplate reader (Molecular Devices).
Oxaloacetate dose response was measured with the Amplite® Fluorimetric Oxaloacetate Assay Kit on a solid black 96-well plate using a Gemini microplate reader (Molecular Devices).
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Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22


Oxaloacetate is an important part of citric acid cycle, where it reacts with Acetyl-CoA to form citrate. It is also involved in gluconeogenesis, urea cycle, glyoxylate cycle, amino acid synthesis, and fatty acid synthesis. The lack of oxaloacetate limits gluconeogenesis and urea cycle function, and can lead to decreased production of energy. Oxaloacetate can be also used as blood glutamate scavengers to provide neuroprotection after traumatic brain injury, expressed both by reduced neuronal loss in the hippocampus and improved neurologic outcomes. Amplite® Fluorimetric Oxaloacetate Assay Kit offers a sensitive assay for quantifying oxaloacetate in biological samples. Oxaloacetate is converted to pyruvate that generates hydrogen peroxide through an enzyme coupled reaction. The production of hydrogen peroxide is monitored with Amplite® Red by a fluorescence microplate reader.


Fluorescence microplate reader

Excitation540 nm
Emission590 nm
Cutoff570 nm
Recommended plateSolid black


Example protocol


Protocol summary

  1. Prepare Oxaloacetate standards or test samples (50 µL)
  2. Add Oxaloacetate working solution (50 µL)
  3. Incubate at room temperature for 30 - 60 minutes
  4. Monitor fluorescence increase at Ex/Em = 540/590 nm (Cutoff = 570 nm)

Important notes
Thaw one vial of each kit component 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. Oxaloacetate standard solution (100 mM):
Add 100 µL of ddH2O into Oxaloacetate Standard (Component E) to make 100 mM Oxaloacetate standard solution.

2. Amplite™ Red substrate stock solution (200X):
Add 50 µL of DMSO (Component F) into Amplite™ Red substrate (Component A) to make 200X Amplite™ Red Substrate stock solution.

3. Oxaloacetate Decarboxcylase (OAC) stock solution (100X):
Add 50 µL of ddH2O into Oxaloacetate Decarboxcylase (Component D) to make 100X Oxaloacetate Decarboxcylase stock solution.


Oxaloacetate standard

For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/13841

Add 10 μL of 100 mM Oxaloacetate standard solution into 990 μL of Assay Buffer (Component C) to get 1 mM Oxaloacetate standard solution. Take 1 mM Oxaloacetate standard solution and perform 1:10 in Assay Buffer (Component C) to make 100 µM Oxaloacetate standard solution (SD7). Take 100 µM Oxaloacetate standard solution (SD7) and perform 1:3 serial dilutions in Assay Buffer (Component C) to get serially diluted Oxaloacetate standards (SD6 - SD1).


1. Add 5mL Assay Buffer (Component C) into one Enzyme Mix1 bottle (Component B1) and mix well.

2. Add 100 μL of ddH2O into one Enzyme Mix2 vial (Component B2) and mix well.

3. Transfer 25 μL of 200X Amplite™ Red stock solution, 50 μL of 100X OAC stock solution, and entire vial (100 μL) of Enzyme Mix2 into the Enzyme Mix1 bottle and mix well to make Oxaloacetate working solution. Note: This Oxaloacetate working solution is not stable, use it promptly, and avoid direct exposure to light.


Table 1. Layout of Oxaloacetate standards and test samples in a solid black 96-well microplate. SD=Oxaloacelate standard (SD1 - SD7, 0.1 to 100 µM), BL=Blank Control, TS=Test Samples. 


Table 2. Reagent composition for each well.

SD1 - SD750 µLSerial Dilutions (0.1 to 100 µM)
BL50 µLAssay Buffer (Component C)
TS50 µLtest sample
  1. Prepare Oxaloacetate standards (SD), 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 Oxaloacetate working solution to each well of Oxaloacetate standard, blank control, and test samples to make the total Oxaloacetate assay volume of 100 µL/well. For a 384-well plate, add 25 µL of Oxaloacetate working solution into each well instead, for a total volume of 50 µL/well. Note: Run the Oxaloacetate assay at pH 6.5 to 7.0.

  3. Incubate the reaction at room temperature for 30 - 60 minutes, protected from light.

  4. Monitor the fluorescence increase with a fluorescence plate reader at Ex/Em = 540/590 nm (Cutoff = 570 nm).



View all 1 citations: Citation Explorer
Metabolic phenotype analysis in patients with pulmonary hypertension
Authors: Ghataorhe, Pavandeep Kaur
Journal: (2016)


View all 17 references: Citation Explorer
Enzymatic assay for D-aspartic acid using D-aspartate oxidase and oxaloacetate decarboxylase
Authors: Kato S, Ikuta T, Hemmi H, Takahashi S, Kera Y, Yoshimura T.
Journal: Biosci Biotechnol Biochem (2012): 2150
Assay of blood and tissue oxaloacetate and alpha-ketoglutarate by isotope dilution gas chromatography-mass spectrometry
Authors: Laplante A, Comte B, Des Rosiers C.
Journal: Anal Biochem (1995): 580
Effect of oxaloacetate and phosphorylation on ATP-citrate lyase activity
Authors: Pentyala SN, Benjamin WB.
Journal: Biochemistry (1995): 10961
Novel oxaloacetate effect on mitochondrial Ca2+ movement
Authors: Leikin YN, Zharova TV, Tjulina OV.
Journal: FEBS Lett (1993): 35
A sensitive multienzymatic assay for the measurement of pyruvate, dihydroxyacetone phosphate, oxaloacetate, and acetoacetate in clear extracts from biological samples
Authors: Arias-Mendoza F, Pina E.
Journal: Prep Biochem (1991): 211
Flow-injection analysis of amino acids and their metabolites by immobilized vitamin B6-dependent enzymes. Sensitive determination of L-aspartate, L-glutamate, 2-oxoglutarate, and oxaloacetate
Authors: Kurkijarvi K, Vierijoki T, Korpela T.
Journal: Ann N Y Acad Sci (1990): 394
Pyruvate dehydrogenase activity in osmotically shocked rat brain mitochondria: stimulation by oxaloacetate
Authors: Haas RH, Thompson G, Morris B, Conright K, Andrews T.
Journal: J Neurochem (1988): 673
Activity of maize leaf phosphoenolpyruvate carboxylase in relation to tautomerization and nonenzymatic decarboxylation of oxaloacetate
Authors: Walker GH, Ku MS, Edwards GE.
Journal: Arch Biochem Biophys (1986): 489
Kinetic mechanism of Escherichia coli isocitrate dehydrogenase and its inhibition by glyoxylate and oxaloacetate
Authors: Nimmo HG., undefined
Journal: Biochem J (1986): 317
The interaction of dithiothreitol and acetyl coenzyme A in a radiochemical assay for rat brain ATP:citrate oxaloacetate lyase
Authors: Simpson J., undefined
Journal: J Neurochem (1981): 100