PT67 cell contamination after transfection - antibody against PT67 (May/11/2007 )

Hi, all

I did transfection of my mesenchymal stem cell (Human derived MSC) culture with retrovirous (created by PT-67 packaging cell line from clonetech) for a while a go, but I got some strange result indicating that vector solution might be contaminated with PT-67 producer cell line, even I filter the vector solution before the transfection.
(so, now, my MSC culture is possibly contaminated with PT-67)

In order to confirm that, we alreday performed PCR for gag, pol, and env genes, and those results were even more confusing, since I did not detect any of those genes in DNA of MSC which potentially contaminated with PT67, but there is a unknown band observed (longer than 1kb, gag, pol, env are less than 700bp)

so I am thinkng that I will try antibody aganinst PT67 or mouse fiblobrast for immunohistochem/immunofluorescent analysis.

so does anyone know a good such antibody specific for mouse fibroblast? Since my MSC is human derived, I am looking something won't cross-react with human.

or if you could give me any suggestion (alternative methods to confirm the contamination), that will be appreciated.

Thanks in advance

Hi,
I think you can try a p24 ELISA to determine the retroviral core antigen.
Good luck.

Andy