Inclusion bodies protein determination - (Jul/18/2008 )

We have our recombinant protein expressed in inclusion bodies we just wash with water and perform solubilization and refolding.

1. can anyone provide me a protocol to determine the total protein content in the inclusion bodies itself.

2. we get our recombinant protein contaminated with host cell potein how to get rid of the host cell proteins.

kindly suggest.

thanks and regards,

Hi

If u solublize ur protein in PBS or similar buffer and u don't have any aggregates, u can just quantified ur

protein by BCA or bradford method and get protein concentration.

Regarding ur other question - there is no clear or simple answer. As i understand u performed Inclusion bodies

purification protocol (its the best). U can use PBS+ 1% Triton x-100 in the washes instead of water (why water?), it helps

to get reed of soluble proteins

U can also can look for various inclusion bodies protocol and make ur own reception of lysis, wash or elution buffers

good luck