Recipe

Luciferin solution

0.2 mM luciferin

15 mM MgSO4

25 mM glycylglycine (pH 7.8)

4 mM EGTA

Just before use, add dithiothreitol from a 1 M stock solution to a final concentration of 2 mM. Approximately 200 μl of luciferin solution is needed per luminometer assay tube. The addition of 1-2 μM CoA to the luciferin solution will produce a more intense and sustained light emission (Wood 1991).

Note:

Luciferin is a generic term for substrates that generate light when acted upon by a luciferase enzyme. The most common luciferase gene in use is derived from the firefly and acts on a particular luciferin substrate. Other luciferase enzymes, such as those from the sea pansy (Renilla reniformis) or certain marine bacteria, use chemically different luciferin substrates. Make sure that the luciferin substrate used in the above buffer matches the luciferase gene incorporated into the expression plasmid.

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  1. doi:10.1101/pdb.rec8836 Cold Spring Harb Protoc 2006: pdb.rec8836-

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