protocols for mouse cells - splenocyte preps, protein extraction (Sep/17/2004 )

Hi there,

Does anyone have a reliable and simple method for culturing mouse splenocytes? I need to stimulate them with mitogen and then I want to look at cytokine production. Any culture medium that works really well?

Also, I am looking for a good protocol for protein extraction, either a commercially available kit or a tried and tested method from someone's lab. I used to do Wblots and the protocol used a protease inhibitor set that was in a pellet that could be dissolved into the lysis buffer each day. Can anyone help with this issue? Thanks very much

Bunster

hi

maybe a look at the "mouse splenocyte prep." topic in the pinned section could give you a clue.

the medium I use for mouse splenocytes is RPMI 1640, 10%FCS, 20mM Hepes, if neccesary 1% Pen/Strep.

mike