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Alanine Colorimetric/Fluorometric Assay Kit

based on 1 citations in multiple journalsAlanine Colorimetric/Fluorometric Assay Kit14.1 4
Catalog #: K652

Product Details

Cat # +Size K652-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The assay can detect serum concentration: ~24-76 µg/ml (~3-9 nmol/10 µl).
Features & Benefits • Simple procedure; takes ~ 40 minutes
• Fast and convenient
• Kit contains all necessary reagents for accurate measurement of alanine levels
Kit Components • Alanine Assay Buffer
• Alanine probe (in DMSO)
• Alanine Converting Enzyme
• Alanine Development Mix
• Alanine Standard (10 µmol)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Alanine is the 2nd most abundant of the 20 proteinogenic amino acids. Nonessential, being available from dietary sources, it plays a key role in the glucose-alanine cycle between tissues and liver. In muscle and other tissues that degrade amino acids, amino groups are pooled as glutamate by transamination. Glutamate then transfers the amino group to pyruvate via alanine aminotransferase, forming alanine and α-ketoglutarate. The alanine is passed into the blood and transported to the liver. A reverse of the alanine aminotransferase reaction takes place in liver. Pyruvate can be used in gluconeogenesis, to form glucose which may return to other tissues through the circulatory system. There appears to be a correlation between alanine levels and and higher blood pressure, energy intake, cholesterol levels, and body mass index. BioVision’s Alanine Assay Kit provides a sensitive detection method of alanine. In the kit, alanine is converted to pyruvate which is specifically detected leading to proportional color (λ=570nm: 0-10 nmol) or fluorescence (Ex/Em 535/587nm: 0-1 nmol) development. Serum concentration: ~24-76 µg/ml (~3-9 nmol/10 µl).

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Will the kit detect D-Alanine?
No, the kit does not detect D-Alanine as the enzyme used is specific for L-Alanine.
Essentially, is pyruvate detected and measured in this assay?
One of the Pyruvate oxidation products reacts with the probe to generate color or fluorescence in this assay. The color or fluorescence is proportional to the amount of Alanine in the sample.
All cells will contain pyruvate. Does that mean there is always background with biological samples with this assay?
If there is endogenous pyruvate, this will generate color in the detection reaction. as mentioned on our datasheet, use background control if high levels of pyruvate are there in the samples. Typically pyruvate concentrations are much smaller than the detectable range of alanine so that background can be subtracted.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically this kit should work with samples from multiple species/sources. Alanine is not species specific.
Can frozen samples be used with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Is it possible to use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
Can a protein assay be used to normalize the loading amount into each well?
Yes, a protein assay can be run with the homogenate/lysate to normalize the loading amount in each well. We suggest using the BCA assay: For serum samples, deproteinization is recommended.
Pang, S. et. al. Regulation of Fasting Fuel Metabolism by Toll-Like Receptor 4. Diabetes, Dec 2010; 59: 3041 - 3048.
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