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After repeated washing captured proteins were eluted from the resin using 10 mM DTT.
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.
The clear supernatant was loaded onto a Ni-chelating column, and bound protein was eluted with 500 mM imidazole.
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