SEAP assay

[Protein Chemistry Facility]

In article <thompsop-2905961612510001 at 203.0.141.239>,
thompsop at burnet.mbcmr.unimelb.edu.au (Phillip R Thompson) wrote:

> I am using Clontech's Great Escape SEAP Genetic reporter system & am
> having a major problem with reproducibility.  Some of my duplicates differ
> by as much as 300%!  The assays are being carried out essentially as
> described in the manual & read in 96-well black plates in a Wallach plate
> reader.  I'm being very careful with pipetting & I don't believe pipetting
> error could result in such large errors anyway.  Any help would be
> appreciated!
> 
I don't know the details of the Clontech protocol, I have used one from
Tropix Inc. . Killing the endogenous alkaline phosphatase by heating at 65
C is essential though. I do this directly in the 96 well plate in a hyb.
oven. 
Hope this helps, 
Breandan Kennedy,
Protein Chemistry Facility,
WAJCSC,
L. Placid, 
NY 12946
Ph  518-523-1268
fax 518-523-4970/1849