Promega Dual Luciferase Assays

Bruno Cenni cenni at cellbio.unige.ch
Mon Oct 18 02:38:55 EST 1999


Do you know the composition of the "passive lysis buffer"? Since they supply only
one bottle of it with their 10'000 reactions kit we'll have to order it
separately... and it's not cheap.

TIA

Bruno

Eric Chace Olivares wrote:

> Hey! We had this same problem in our laboratory, and I'll tell you how it
> was fixed.  The incompatibility with the BioRad protein assay kit comes
> from the levels of glycerol and DTT present in the lysis buffer.  By
> remaking the lysis buffer with reduced amounts of these, we were able to
> obtain results using the BioRAD DC-protein assay (detergent compatible).
>
> If you want the exact recipe for the new lysis buffer, you can email our
> tech at ebaba(at)stanford.edu.
>
> Hopefully this helps.
>
> -=Eric
>
> Windows Manager (WINMAN at leeds.ac.uk) wrote:
> : We wish to compare the activity of a promoter construct in various
> : different cell lines which have both different viability in culture and
> : different transfection efficiencies.  The Renilla control appears to correct
> : for transfection efficiency but not the different viabilities.  We have tried
> : the BioRAD protein assay but this appears to be incompatible with the Promega
> : Passive Lysis buffer.
> :
> : Has any one found a way to correct this system ?  How else could we correct
> : this variability ?
> :
> : Thanks
>
> --
> Eric C. Olivares
> Department of Genetics
> Stanford University School of Medicine
> olivares at stanford.edu
> http://www.stanford.edu/~olivares/




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