NEB/Luciferase Cell Lysis Buffer/B3321S/25 ml

Description:

Luciferase Cell Lysis Buffer (LCLB) is a proprietary formulation developed to produce mammalian cell lysates for reporter assays. This lysis buffer is compatible with reagents for assaying the activity of Gaussia as well as other luciferases (e.g. Renilla & Firefly) and β-galactosidase (Figure 1A-D).

Figure 1: LCLB compatibility with commonly used reporter assay systems

Notes:

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).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.