SEAP

[Dan Johnson]

In article <olems-1103961959240001 at vi05.imb.fm.uit.no>,
olems at petrus.fagmed.uit.no (ole morten Seternes) wrote:

> We have just bought the the SEAP reporter system from Clontech. We are
> currently using CAT-assays in our transient transfection studies, but the
> Secreted alkaline phosphatase system loocked much easier. But we are
> unable to detect any  SEAP activety after transfection of NIH-3T3 cells.
> We have used both calsium phosphate mediated and lipofectamin mediated
> transfection methods. I wonder if there are anybody that have used this
> reporter gene system with success?
> 
> 
> Ole Morten Seternes

	   First, a clarification.  Are you using a colorimetric assay for
alk.phos., or luminescent?  I have gotten the colorimetric method to work
minimally with COS cells, but in muscle cells the limit of detection is too
low, although DNA is getting in to the cells.  You might try a luminometric
assay instead.  Also, I found Clontech's particular  constructs are not
very active, so you may have to go to way more DNA than they specify.  Good
luck. 

Dan Johnson