I tried to do a Slot Blot and was not successful and melatonin is only .23 Kda so it would probably run off a western blot. Has anyone had success with other methods?
Hello Hannah
You can try chemiluminescence method which is based on the emission of chemiluminescence when melatonin reacts with hydrogen peroxide and acetonitrile under alkaline conditions.
You can also try determining melatonin spectrophotometrically after reaction with potassium permanganate and formaldehyde in acid medium.
For details you can refer to the link below:
https://ijpsr.com/bft-article/melatonin-a-review-on-the-lesser-known-potential- nutraceutical/?view=fulltext
Regards.
ELISA:
https://www.biocompare.com/pfu/110627/soids/2-4313/Assay_Kit/ELISA_Melatonin
Have you considered doing a DOT blot instead?
Rather than run the sample on a gel and transfer, you just prep some membrane and sport the sample onto it. Once dried, block and run as usual.
It's normal to do serial dilutions of the sample and spot across the membrane.
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