Insect cell (SF9) flow

[Nina7]

I am looking for a protocol for insect cell flow. Actually I did find it, but I do need to treat my cell with (Brefeldin) BrA or Monesin and wanted to know if they can be used in insect cells and what concentration.

Thanks a lot!

[sivaprakash]

hi
I am purifying a protein which is a kinase family from E.coli overexpressed cells.I am not getting the protein in cytoplasm (ie, soluble fraction) after IPTG induction.hence, I have tried with reduing temperature (28 C) and also reduced the IPTG concentration (0.5 mM). But still it goes to inclusion bodies.
Experienced people in this field can help me

Thanks
Siva

[sivaprakash]

Hi
I am trying to purify the soluble protein overexpressed in E.coli. Which is a kinase family. Intially, I have tried with 37C and 1mM IPTG not worked it went to inclusion bodies. And I reduced the ITG concentration to 0.5 mM and temperature (28C). But still I am not getting recombinat protein in cytoplasmic fraction.

How to overcome this problem

Siva