protein expression in EL-4 stable transfectants

Soo Jung Yoon j2w19may at chollian.net
Wed Jun 30 05:35:22 EST 1999


Hi, there,

I'm trying to establish stable EL-4 transfectants by Lipofectamine
procedure (they're going to be used as target cells in a CTL assay), and
I've got growing clones after G-418 selection for 2 wks at 500 ug/ml.
Now, what would be the best procedure to confirm insert-protein
(cytoplasmic) expression? I've tried dot blot of whole protein fraction
once, but there were no difference with the mock sample.

Which one of dot blot, Western blot, or Immunoprecipitation would be
better? Should I use isotopes if only trace amount of protein exists?

Thanks in advance.

Best Regards,
Sujeong





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