I am using TALON® Metal Affinity Resins for purification but after use, it ones colour of beads changes from faint pink to dark pink colour.
For cleaning the column I used 20mM MES with 0.3M NaCl buffer (as recommended in company protocol).
Result of it doesn’t have an affinity to his tag
A change of color must happened if new molecules are participating in the coordination process with the metal ion. Also you can expect remotion of some metal ions. Anyway why are you cleaning it? Do you have protein precipitation?
The company protocol mention that with 6M GuHCl it’s color becomes dark pink color but after washing with 10-20 col of water color remains unchanged but
after that i washed with 20mM MES buffer( pH 4) then also beads are dark pink color
The color should be the original after you pass your adsorption buffer, if this does not happen some molecules are still adsorbed. Remove the ions with a 50 mM EDTA solution and load new ions.
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