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Step  2, First Ammonium Sulfate  Fractionation
Solid ammonium sulfate was added  to  the cell-free extract  to give  56% saturation  (at0¡æ). The pH was maintained at 6.9 with 2 N KOH. After stirring for
1  h  and removal of  the precipitate by centrifugation, the  supernatant
solution was further  saturated with ammonium  sulfate  to give 72%
saturation  (0¡ãC). After  I h, the suspension was centrifuged, and  the
pellet was dissolved in Buffer A containing 1.25 M  ammonium sulfate
and 0.03%  sodium azide.
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