INDIGO-Ni Regeneration Protocol

 

INDIGO based agarose resins and MagBeads should be washed & regenerated at least after every 5th use. Though we recommend to perform this regeneration protocol after each use, if possible.
Note: The INDIGO ligand cannot be stripped with EDTA
 
Note: "CV" refers to column volume, I.e. for 1 ml column bed volume use 10 CV = 10 ml of buffer
  1. 10 CV H2O
  2. 10 CV 500 mM NaOH
  3. 10 CV H2O
  4. 10 CV Neutralization buffer (150 mM sodium chloride; 200 mM Na2HPO4, pH 7.0)
  5. 10 CV H2O
  6. 10 CV 20% (v/v) Ethanol, 10 mM sodium acetate, pH 6.50*
Note: 20% (v/v) Ethanol, 10 mM sodium acetate, pH 6.50, is the recommended storage buffer for the column and resin.
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