Virtualab Protocols (Chang Bioscience)
1. Inclusion body is precipitated, insoluble proteins in bacteria.
sure your protein is in inclusion body. Membrane-bound protein may also
be insoluble without detergent, but can be solublized by including low
concentration (<1%) of SDS or Triton X-100.
3. Attempts to avoid inclusion body include growing bacteria at different
temperature and using different bacteria strains and culture media.
4. Inclusion bodies are frequently solublized with a buffer
that contains 8 M urea or 6 M guanidine hydrochloride.
5. Refolding may be achieved by:
-Quick dilution, e.g., dilution of the proteins in
8 M urea to 0.4 M urea.
-Refold on column, e.g., changing buffer from
high urea or guanidine hydrochloride to no urea or guanidine hydrochloride
while protein is being purified by ion exchange column or affinity column.
-Slow dialysis against a buffer without
-Refolding in a refolding buffer.