I am working in a college lab. We can't use expensive enzymes. Please suggest an easier method for extraction of melanin from human hair.
Hi Noble
you can use different methods, depending of your source material, with physically harsh conditions:
http://www.biochemj.org/content/15/1/118
http://www.academicjournals.org/journal/AJMR/article-full-text-pdf/B741D6F22127
http://ebm.sagepub.com/content/25/8/635.extract
There is a possibilty to completely digest Keratin enzymatically with cheap technical Proteases, they are easy to get in high activities and will not outlimit your budget.
You need to add a SH-Bond reducing agent, ß-Mercaptoethanol, Thioglycolic Acid etc.
After you can extract the Keratin more easily. This "Organosolv-Keratin" is more native, so you can choose depending on if you only want to de-ink hair or really want to isolated native Keratin.
Good Luck
Hi Noble,
I have successfully extracted melanin from mouse hair using a simple assay. I then went on to measure the absorbance at 500 nm, which serves as an indicator of the total eumelanin and pheomelanin.
The method is from this article:
"Advanced Chemical Methods in Melanin Determination" (Wakamatsu et al, 2002)
http://onlinelibrary.wiley.com/doi/10.1034/j.1600-0749.2002.02017.x/abstract
DOI: 10.1034/j.1600-0749.2002.02017.x
Note that this article states that this method has been successfully used to dissolve mouse hair, sheep wool, and human hair samples.
This is how I performed the assay using shaved mouse hair:
Dorsal hairs of mice at P50 or P100 were shaved and 1 mg was dissolved overnight in 1 mL of hot (65°C) Soluene-350 (PerkinElmer) and 10% water. Triplicate 150 μL aliquots for each mouse hair sample were analyzed for absorbance at 492 nm as previously described to generate an average absorbance value.
Hope this helps & good luck
-Elyse
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