Immobilization of K lactis galactosidase Onto Thiol Reactive Supports

2.3.1. Activity Determination

1. Activity buffer: 20 mM potassium phosphate buffer, pH 7.0, containing 2 mM magnesium chloride, 0.1 M, KCl.

2.3.2. Protein Determination

1. Coomassie Plus protein assay reagent (Pierce, Rockford, IL).

2.3.3. Immobilization of the Enzyme

1. Thiolsulfonate-agarose (TS-gel) and thiolsulfinate-agarose (TSI-gel) prepared as described in Subheadings 3.1.3. and 3.1.4. respectively.

2. Immobilization buffer: 0.1 Mpotassium phosphate buffer, pH 7.0.

3. 0.1 Mpotassium phosphate buffer, pH 7.0, 0.5 MNaCl .

4. P-Galactosidase (P-D-galactoside galactohydrolase; EC 3.2.1.23) from Gist-Brocades (Gif-sur-Yvette, France).

2.3.4. Blocking Excess of Active Groups 1. GSH (Sigma)

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