Dear Bob,
Have you made any attempt to grow your E. coli (that are recombinantly
expressing the protein you're studying) in different media to eliminate the
presence of inclusion bodies? In the reference "Overcoming Inclusion Body
Formation in a High-level Expression System" (J. T. Moore et al., (1993)
Protein Expression Purif., pp 160-163), the authors describe an "enriched"
growth medium they tried/developed to over-express their enzyme (using the
pET system from Novagen) in an active form with good results. Previous to
doing this, their enzyme was expressed in an inactive form in inclusion
bodies. Of course, if all you need your protein for is to generate
polyclonals, activity may not be an issue here. Just the same, this might
be an alternative approach you might want to consider.
Best of luck,
Bob Keefe
Genomics Core Facility
Wadsworth Center/NYS DOH
At 01:43 PM 9/13/2000 -0400, you wrote:
>Hi,
>
>Need a good way to prepare the protein from the inclusion body of the E.
>coli for raising polyclonal antibody. Thanks.
>
>Bob
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