I have been trying to check the levels of ROS and NO in RAW cells after treating with my recombinant protein. Every time I do the experiment, I get high levels even in Proteinase treated group. I treat the protein with polymyxin beads to remove LPS. I do not get such high readings in the cells that are treated with active protein. Can anyone explain?
I have run the proteinase treated sample on the SDS gel to check if the degradation had happened. I do not see any band.
Chandra Somasundaram
How do you treat the cells with recombinant protein? did you transfect the cells with recombinant DNA?
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Kala Jyothi
I purify, concentrate and dialyse the protein sample; then treat with polymyxin beads to remove LPS and treat the cells with different concentrations.
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