Hello. I hope to get an explanation why I should trash my 6-month long effort. I cloned a mRNA from a patient into pMX retroviral vector (as either C or N-terminal Flag fusion protein). When I transiently transfect HEK293 with this construct I do not detect my protein with the Flag antibody. The cells are transfected since the retorviral vector has a GFP (translated from IRES) and the cells are green (at least 30% of them). I also cloned this mRNA into pcDNA3.1+ vector and the expression is beautiful. What am I doing wrong? Can the protein form retroviral vecotrs be deteted from transient transfections?
I am eager to learn so please teach me. Thanks
Edited by KristinaZ, 24 September 2013 - 07:04 AM.