How to prepare home-made luciferase substrate

[martin LEACH]

Hi...I dug this recipe out of a colleague's thesis..
I havent tried it yet though...

20 mM Tricine
1.07 mM (MgCO3)4Mg(OH)2.5H2O
2.67 mM MgSO4
0.1 mM EDTA
33.3 mM DTT
270 uM Coenzyme A
470 uM Luciferin
530 uM ATP

pH 7.8

this may be the original recipe and not the proprietry Promega one...

in my experience I couldnt get luciferin to dissolve in water I used a
luciferin potassium salt...

hope this helps 

Martin 

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Dominic Voon (dvoon at uniwa.uwa.edu.au) wrote:
: Hi all,
: 	The Promega Luciferase assay kit that we purchased is running
: out of  luciferase substrate (luciferin, CoA and a mysterious
: competitive inhibitor of luciferase), and we don't want to splash out
: big bucks for another. Would someone be so kind as to tell me (and the
: newsgroup !!) how to make my own luciferase assay substrate ? Any
: information will be much appreciated. If possible, send a copy of your
: followup post to me via email at dvoon at cyllene.uwa.edu.au.


: Many thanks in advance :-)

: dominic