I used Lipo3000 to transfect the Htt-16Q and Htt-148Q plamids in HEK293 cells and PC12, SHSY5Y cell lines. The results in HEK293 are very good. But, I cant see any Htt expression in transfected PC12 or SHSY5Y cells. Can anybody do the similar experiments and give some tips? In addtion, after transfecting the Htt-148Q plamid, I never see the cell death even after 72hours, is there something wrong?
Hello Sun. I used Lipo2000 to transfect PC12 with mEGFP-Lifeact-7 plasmid. The results are good. I don't know if there is a different between use Lipo3000 or Lipo2000, if still not working I can give to you my protocol to transfect.
Want to infect PC12 cells to look at changes in general expression. Is it vest to use cells in suspension or adherent to the dish?
Hi Sun,
I have recently tried to transfect Htt-18Q and Htt-148Q plasmids in SHSY5Y cells (in a 96 well plate) and am having difficulties in measuring cytotoxicity. I don't see any cell death with the 148Q plasmid. I have been trying to figure out the problem. Do you have any suggestions? Did you change any protocols and observe the cytotoxicity/cell death with the 148Q plasmid.
Thank you.
Khushboo.
Hi! Did you find the way to transfect efficiently PC12 and SH ? I'm having the same problems, and I don't get better results thant 40% in SH and 15% in PC12 using electroporation with AMAXA Nucleofector.
Thank you very much,
Best,
Ana.
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