Hi, everyone, I am recently working on the baculovirus protein expression system. The bacmid I used is from our lab stock. There is no problem with the insert. Sf9 cells infected by a P1 virus for 4 days. The cells was larger and lysis compare to control cells. And some dead cell clusters floating in the culture medium. Is that normal?
Then I collected the cells followed by Western Blotting check. But there was no band could be detected.
Any suggestion will be appreciated.
Thank you very much.
Hi,
As the infection period is prolonged, the cells can lysis too much. But, it is normal that Sf9 cells infected with bacmid are getting larger, can be lysis and float in the culture medium.
For western blot analysis you can wash your pellet with 1X PBS included protease inhibitor coctail after collecting cells. In addition, if there is no expected band, you can use culture medium with FBS to reduce degredation of desired protein.
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