To isolate the Tyrosinase enzyme we have eliminated melanin using sephadex. Now melanin is on sephadex. Does any one have an experience regarding the step that could release the remaining melanin from sephadex (elution)? Thanks!
Find a solvent that is capable of dissolving the melanin and is also compatible with Sephadex. Apparently, melanin is soluble in aqueous sodium hydroxide. Sephadex can be washed with 0.2 M NaOH, so try that. The recommended method for cleaning Sephadex columns is on page 7 of this manual..