EnzyChrom™ L-Carnitine Assay Kit

Protocol SDS

Application

• For determination of L-Carnitine in urine, serum, plasma, cell lysates, tissue lysates and food products (e.g. milk).

Key Features

• Sensitive. Uses 10 µL samples. Detection range: colorimetric assay 12 – 1000 µM, fluorimetric assay 1 – 100 µM L-Carnitine.
• Convenient. Room temperature “mix-incubate-read” procedure can be readily automated for high-throughput assay of thousands of samples per day.

Method

• FL530/585nm or OD570nm

Samples

• Urine, serum, plasma, cell lysates, tissue lysates and food products (e.g. milk).

Species

• All

Procedures

• 30 min

Size

• 100 tests in 96-well plate

Detection Limit

• 1 µM

Shelf Life

• 6 months

More Details

L-CARNITINE is found in most mammals, plants, and some bacteria. L-Carnitine’s most crucial role is in the transport of long-chain fatty acids from the cytosol into the mitochondria for fatty acid oxidation. BioAssay Systems’ method provides a simple and high-throughput assay for measuring L-Carnitine. In this assay, a multistep reaction produces H₂O₂ which reacts with a specific dye to form a pink colored product. The optical density at 570nm or fluorescence intensity at λex/em = 530/585 nm is directly proportional to the L-Carnitine concentration in the sample.

Why does the cloudy Substrate Mix not interfere with the assay?

The substrate mix components are at the edge of solubility in DMSO, but once diluted in assay buffer, they are completely soluble. Unfortunately, there isn’t a universal solvent that allows for a concentrated solution for all three substrates.

Can this kit be used to study the kinetics of Carnitine Acetyltransferase?

No. Because one of the two substrates for Carnitine Acetyltransferase is included in the Substrate Mix and not separate, it would complicate the downstream reactions to vary the concentration of the substrate mix.

I am having difficulty detecting low levels of L-Carnitine in a biological sample. Is there anything I can do to facilitate the quantification?

You can try using an internal standard with biological matrix you are testing. To do this, you will employ three different wells, one with Sample, one with Sample plus Internal Standard, and one that will be the Sample Blank. The Sample well and the Sample Blank well each receive 10 µL of sample while the Sample plus Internal Standard well receives 10 µL of Sample plus 0.25 µL of 1 mM L-Carnitine. The Sample well and the Sample plus Internal Standard well will receive 90 µL of Working Reagent while the Sample Blank well receives 90 µL of Sample Blank Working Reagent, which has all the components of the Working Reagent minus Enzyme A. See the protocol for the details of the downstream analysis.

For more detailed product information and questions, please feel free to email or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to Technical Support.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.