Hi Pouya,

Thank you for the suggestions. I went through those articles before I post this question. All of them have used radio labeled fatty acids.

I am looking for a assay or a kit can be used without radio labeling.

Abcam provides a kit to detect Medium-chain-acyl-CoA dehydrogenase (ab129734) in both cells and tissue extracts. I'm curious to know if there any other method available without radio labeling.

Thanks,

Susara

good I follow the answers

Hello Guys..

I found a less expensive kit to detect fatty acid oxidation. The assay is based on the oxidation of octanoyl-CoA, which is coupled to NADH-dependent reduction of INT to INT-formazan. The formazan product exhibits an absorption maximum at 492 nm, allowing for sensitive measurement of FAO activity present in cell/tissue extracts.

For more information-

http://www.bmrservice.com/FAO.html

Good luck !

beta-oxidation of fatty acids measured by quantitation of 3H-labeled H20 generated at the acyl-CoA dehydrogenase step.

Hello Susara,

Finally did you obtein good results with the FAO assay kit from BMR? I'm really interested in using it, I found this kit has a better and easier protocol than anothers FAO assay kits. Thank you very much!

Hi Emma,

Yes. The assay results were aligning with our western blot results. This kit is very straight forward. I recommend it :)

Good Luck..!

Susara

Dear Susara Madduma Hewage ,

Following your recomendation I also bought the BMR kit to check B-oxidation in skeletal muscle.

I am following the protocol with gastrocnemius muscle and I checked different concentrations, my problem is that I got the same colour and absorbation in the control well than in the substrate well....Has this happened to you? Could you give me some advice with your eperience?

Thanks you!

Dear Erica,

Apologies for the late reply. No we didn't see such a problem with our samples. Our substrate values were around 0.1440 while the sample values were varied from 0.25 to 0.45.

I feel like your samples are too much diluted.

Good Luck,

Susara