EnzyFluo™ Fatty Acyl-CoA Assay Kit

Protocol SDS

Application

• For quantitative determination of fatty acyl-CoAs in tissue and cell lysates.

Key Features

• Sensitive and accurate. Use as little as 10 µL samples. Linear detection range in 96-well plate: 0.3 to 100 µM fatty acyl-CoA. Internal standard method minimizes potential sample matrix interferences.
• Fast and convenient. Room temperature assay with a 40 min incubation. No 37°C incubator is needed.
• High-throughput. Homogeneous “mix-incubate-measure” type assay. Can be readily automated to assay thousands of samples per day.

Method

• FL530/585nm

Samples

• Tissue and cell lysates

Species

• All

Procedures

• 40 min

Size

• 100 tests in 96-well plate

Detection Limit

• 0.3 µM

Shelf Life

• 6 months

More Details

Fatty Acyl-CoAs are a combination of Coenzyme A covalently bound to long chain fatty acids. Fatty acyl-CoAs serve as both a precursor for triglyceride and phospholipids synthesis, and a product of lipid catabolism. Catabolically, they facilitate the transport of fatty acids to the mitochondria for shuttling via the carnitine shuttle and subsequent β-oxidation of the fatty acid. Conversion of fatty acids to fatty acyl-CoAs is necessary for generation of ATP during periods of high energy demand. Elevated fatty acyl-CoA levels may indicate dysregulation of lipid metabolism such as due to insulin resistance. Certain genetic disorders may also affect fatty acyl-CoA levels. BioAssay Systems’ fluorimetric fatty acyl-CoA assay works using a combination of enzymes that utilize fatty acyl-CoA as a substrate, which is coupled to form a fluorescent product. The resulting fluorescence intensity at λ_exc/em = 530/585 nm is linear to the fatty acyl-CoA concentration in the sample.

How should I store my samples?

Fatty acyl-CoAs are not stable and samples should ideally be tested the same day. Otherwise, snap freeze the samples in liquid nitrogen or freeze at -80°C for up to one week.

Can this kit be used with serum or plasma samples?

In our testing, the concentration of fatty acyl-CoAs was too low to detect in these samples.

Can I use a different lysis buffer?

We suggest using a lysis buffer that is pH 7.4 and is 0.5% – 5.0% triton X-100 buffer to dissolve fatty acyl-CoAs.

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.