I have tried several different reagents and monitored from 4 to 72 hours, I am not seeing toxicity, but I am not getting siRNA into the cell, no detectable GFP with contro and no knock down with experimental
Hello
Look at links here will help you
Good Luck
http://search.proquest.com/openview/8570be9c210586117eba870f16feaa07/1?pq-origsite=gscholar&cbl=55345
http://www.jneurosci.org/content/26/50/12904/suppl/DC1
Thank you for your response, I've seen these papers and many more with successful transfections in BV2 cells. While I have been able to transfect other cell lines I have not been able to get it to work in the BV2s . I thought perhaps there was a "secret" that never makes into the manuscripts.
A while ago, I also had difficulty in transfecting BV2 cells, and tried many different transfecting reagents that easily work for other cell types. My BV2 cells either did not get transfected or got transfected with low efficiency but the cells were activated (problem with immune cells)! In the end, I got it work using Nucleofection (Lonza), efficiency is around 50-60% and it is gentle on cells. Maybe you could look into it if someone or your lab has the machine.
http://bio.lonza.com/sirna-concentration.html
Thanks, I may try that. I actually got them to transfect on chamber slides but not on plastic. Not sure what that is all about.
Diane, can you please share the protocol that has been working for you on chamber slides? We are also having some problems transfecting BV2 cells. We have tried lipofectamine 2000, and glial-mag but with very low efficiency. There is some report that TransIT®-LT1 from Myrus might work, but we have not tried it.
Article Translocator protein (18 kDa) (TSPO) is expressed in reactiv...
Hi Arie, I have used lipofectamine RNAiMax, Xtreme gene siRNA, and siRNA transfection reagent from santa cruz. All of them gave me about 30% transfection efficiency. I had to use 300 nM siRNA and the high end of the suggested transfection reagent concentration. I have since changed to a lentiviral particle transduction.
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