Luciferase Cell Lysis Buffer

1. Depending on the cell type, some cells may need larger volume and longer incubation time for sufficient lysis. One can determine whether or not the lysis is sufficient by assaying the activity of intracellular Gaussia luciferase using the pCMV-GLuc Control Plasmid (NEB #N8081). Although the Gaussia luciferase produced from pCMV-GLuc is secreted into the culture medium, at a given time point, 5-10% of the total activity should reside in the lysate (Figure 1A).
2. Normally, there is no need to remove the cell debris by centrifugation before assaying Gaussia luciferase activity. If additional assays are to be performed (e.g. protein concentration, SDS-PAGE, etc.), it is best to centrifuge the cell lysate to remove cell debris.

1. Luciferase Cell Lysis Buffer

• Ultrasensitive Secreted Luciferases B3321

1. What is the composition of the Luciferase Cell Lysis Buffer?
2. Can a lysis buffer other than the Luciferase Cell Lysis Buffer be used to make cell lysates for assaying GLuc (or CLuc) acitivity?
3. Can luciferases other than GLuc and CLuc be assayed from cell lysates made with the Luciferase Cell Lysis Buffer?
4. Are there differences between cell lines on how fast they can be lysed with the Luciferase Cell Lysis Buffer?
5. Do I need to remove cell debris in the cell lysate before assaying for GLuc or CLuc activity?

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• Luciferase Cell Lysis Buffer

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