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After extensive washing of the unbound protein with 40 mM imidazole, the GHS protein was eluted with 250 mM imidazole.
Bound material was then eluted with 0.2 M methyl mannoside in the same buffer.
A radioactive compound identified as p-coumaroyl ß-glucosyl ester was eluted and stored frozen.
After repeated washing captured proteins were eluted from the resin using 10 mM DTT.
The clear supernatant was loaded onto a Ni-chelating column, and bound protein was eluted with 500 mM imidazole.
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