I want to do one differentiation assay in pre adipocytes 3T3 L1 cell lines and I have tried the transfection with calcium phosphate method and X-fect clontech reagent, but I m unable to get transfection.. so please help me rectify this problem
3T3-L1 are very difficult to transfect, but we have had some success with Lipofectamine 3000 using their standard transfection protocol.
PolyJet PEI works, as does Mirus' Trans-IT x2. However, with cationic lipid based reagents it is hard to do better than 15-30% transfection efficiency in this cell line. Higher efficiencies are possible using AMAXA's nucleofection system.
Hi Mamta,
Maybe is late but you can try it with DeliverX Plus Transfection kit (Affimetrix), the transfection rate can be quite good.
Have you already get it? Good luck!
Xtreme Hp DNA transfection reagent from roche with their recommended protocol has worked very good for me. But I had to optimize the seeding density and DNA ug/well to get good efficiency in co-transfection experiments.
If you are using this, you can retain the transfection mix after 6 hours of transfection and add regular growth media on to it.
https://lifescience.roche.com/wcsstore/RASCatalogAssetStore/Articles/Protocol_X-tremeGENE%20HP_3T3-L1.pdf
Yeah 3T3 L1 is pretty tough, given that it's an embryonic fibroblast cell line. I know some have gotten 70% efficiency with siRNA transfection using specialized reagents (https://altogen.com/product/3t3-l1-transfection-reagent-emb-fibroblast-cells-cl-173/), but it also depends on what you are transfecting; larger plasmids will have far lower efficiency.
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