transfection and expression in human PBMC

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mbalan
mbalan's picture
transfection and expression in human PBMC

HI, I am a Grad student at UMDNJ NJ USA..we have been trying to transfect human PBMC and follow the expression with egfp.The positive control was pmaxGFP supplied with the Amaxa transfection kit.we have tried different possibilities but no expression.. we have changed the promoter of our constructs to CMV and we have an IRES in the construct.. the size of the plasmid is about 9kb. please suggest about transfection in human PBMC. thanks, BASKAR

saswati1
saswati1's picture
Why don't you check the

Why don't you check the following website.

http://www.biocompare.com/techspotlight/180/Opening-The-Door-To-Transfection.html

They have FuGENE 6 Transfection Reagent - Roche Applied Science, that might be of your interest.
Hope it helps

lstern
lstern's picture
Have you tried to stimulate

Have you tried to stimulate the PBMCs first with 4ug/ml PHA for 48 hrs or 1ug/ml each CD3/CD28 prior to tranfection. This works pretty well