Fab Fragments Rabbit

Dialyze F(ab02 fragments against Soln. A. Add 2-mercaptoedianol to a final concentration of 0.2 M (15 pl/ml). Incubate at RT for 10 min, cool in an ice bath, and add iodoacetamide to a final concentration of 0.3 M (55.5 mg/ml). Incubate on ice for 1 h and dialyze against PBS or desalt on a SephadexG-25 column, equilibrated with PBS (Fab fragments in the void volume). 

Add 1/20 volume Soln. C, 1/20 volume Soln. D and 10 pg papain/mg IgG to Soln. B. Incubate at 37 °C for 8-12 h, and then add 13.8 mg per milliliter assay volume of iodoacetamide. Allow to react at RT for 30 min, then dialyze against PBS or separate on a Protein-A column (Fab fragments in die passage). 

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