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Transfection of SH-SY5Y cells

SH-SY5Y neurocell transfection transient luciferase

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#1 Voneyvoney

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Posted 17 September 2014 - 02:26 AM

Hi All Anyone has experience in transient transfection in SH-SY5Y cells? I will need to perform transient transfection for downstream luciferase assay analysis. I have tried seeding the cells to 70% confluence following by standard lipofectamine2000 protocol. But there are only about 15% of cells expressing RFP 4 days after transfection. I'm welcoming any suggestions. Thank you!!!

#2 bob1

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Posted 17 September 2014 - 02:01 PM

Transient transfection using lipid-based techniques like LF2000 are unlikely to get you more than 30% transfection anyway, even in the easiest to transfect lines (e.g. 293, HeLa). If you need a higher transfection rate, consider using electroporation methods.



#3 Voneyvoney

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Posted 21 September 2014 - 06:11 PM

Thanks Bob! I have tried electroporation method with 10 ug DNA/ 100 mm petri dish, the RFP expression was very low (<10%) too.  

Thus far, beside the Lipo2000, I have also tried Genefactor and Novafector, all these transfection doesn't seems to work for SH-SY5Y cells even though the ratio and so had been change.

Anyone have experience transfecting SH-SY5Y using calcium phosphate method?unsure.png



#4 rkay447

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Posted 22 September 2014 - 12:19 PM

Some lines just don't transfect well.  That's all there is to it.  You might want to try the lipofectamine plus system or think about constructing an lentivirus.



#5 bob1

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Posted 23 September 2014 - 01:10 AM

Calcium phosphate systems tend to give poorer transfection than lipid based too.







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