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Luciferase Cell Lysis Buffer |
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Prices are in US dollars and valid only for US orders.
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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. NIH3T3 cells were transfected either with (A) pCMV-GLuc Control Plasmid (NEB #N8081S), (B) a lacZ-expressing vector, (C) a Renilla-expressing vector or (D) a Firefly-expressing vector, using TransPass D2 Transfection Reagent (NEB #M2554S). The supernatants from CMV-GLuc-transfected cells were saved for the assay at 24 h after transfection. All transfected cells were then washed once with PBS (pH 7.4) and lysed with either Luciferase Cell Lysis Buffer or commercially available lysis buffers recommended for each reporter system, respectively. Cell lysates (20 µL out of 100 µL total lysate per sample) were analyzed for GLuc or Renilla luciferase activity using the GLuc Assay Kit (NEB #E3300S/L), β-galactosidase activity using Galacto-Light System (ABI), and Firefly luciferase activity using Luciferase Assay System (Promega). Gaussia luciferase activity was also assayed from the supernatants (20 µL out of 500 µL growth medium of CMV-GLuc-transfected cells per sample).
Reagents Supplied:
Luciferase Cell Lysis Buffer
1X Luciferase Cell Lysis Buffer:
1 X Proprietary formulation
Storage Conditions
Storage Temperature:
4°C
Notes
Usage 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.
Companion Products
Gaussia Luciferase Assay Kit
pCMV-GLuc Control Plasmid
pGLuc-Basic Vector
TransPass™ COS/293 Transfection Reagent
TransPass™ D2 Transfection Reagent
TransPass™ HeLa Transfection Reagent
TransPass™ HUVEC Transfection Reagent
TransPass™ R1 Transfection Reagent
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