[quote name='Clare' date='May 21 2009, 01:03 PM' post='24855']
Have you tried doing a spin infection? I found this was better than just culturing in polybrene/virus. It gave me a much better % of GFP+ cells.
In some of my experiments I did stimulate them as I was interested in what effect overexpressing a particular gene would have on antibody secretion. In these experiments I used resting B cells.
What exactly do you want to study in your B cells?
Clare'B cells are brilliant, T cells are terrible!'
Thanks Claire. I did a spin infection once but on sterile FACS tube. Incubating the tube overnight and then transfer the cell suspension to 12-well plate but this didnt work. I will try the spin infectin on the plate itself as you suggested and hopefully this will solve the problem.
I am trying to do an in-vitro knockout on immune cells using lentivirus. My gene of interest already flanked with two loxP sites in all mice cells so when I add the viral particle carying Cre recombinase this will result in deletion of my target gene. After achieving this knockout system I am going to test the effects of deletion of this gene on the immune cells starting from B-cells cuz they are brilliant as you said
Edited by medo, 21 May 2009 - 09:13 AM.