Hi, All:
Although it is off-topic, definitely I think I can get some help here
because we crystallographers are dealing with protein purification almost every
day.
My protein was expressed in E.coli as a soluble octamer with or without
his-tag revealed by gel-filtration. For the his-tagged protein, the final
product is an octamer after Ni-column and gel-fil. However, purification of the
non-tagged protein results in a tetramer because of a partially denatured
condition.
When I tested the enzymatic activity , it turns out the tetramer was
active although both of them can be crystallized. Now I am wonderring whether
its native form in human is an octamer or tetramer.
I am planning to purify a little protein from human cells to verify its
native form.
Can anybody recommend some protocols?
Thanks a million,
Jerry McCully
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