I'm not sure about your second question, but FuGENE HD from Promega works well with RAW 264.7 cells. They have a standard protocol that can be found in their protocol database: http://www.promega.com/techserv/tools/FugeneHdTool/

Thanks Jordan

Dear,

at first, please forgive the fact that, as a specialist of transfection, I will only answer to the firts part of your question,

OZ Biosciences developped transfection reagents since almost 15 years (please refer to the link below) and we have reagents that are really efficient for Raw 264.7 transfection.

Would you like to try it, please do not hesitate to contact me via ResearchGate or directly at [email protected]

best regards,

Cedric

http://www.ozbiosciences.com/

In order to get information about homodimerisation using e.g. co-IP you do not need to transfect a plasmid DNA vector containing two gene inserts for the two deferentially tagged versions of the same gene. Simply co-transfect two independent plasmids encoding the respective genes. Both pDNA constructs will be encapsulated into the transfection complexes equally well (actually many genes can be co-transfected without problem). We have also developed our own transfection reagent (ScreenFect A) in our lab at KIT and we actually sell it through the company Incella. It is attractively priced and we can send you test samples if you like. Good luck!

hi thanks Gary but two independent plasmids should be same or can we different?I will like to test your reagent. Let me know the procedure.

they can be the same or different expression vectors, does not matter. Main point is they are both expressed in mammalian cells to similar extent (e.g. expression of tagged gene insert driven by the CMV promoter). So if you already have two different mammalian expression constructs of your gene (harbouring different tags) you can use them straight away in your transfection. Please contact Marvin Krebs ([email protected]) to arrange for us or one of our distributors to send you a test sample of ScreenFect A.