Transfected 293T cells seeded in 12-well plates were harvested using 350 ml of Reaction Lysis Buffer from the Luciferase Assay System (Promega) per well. Cell lysates were diluted 1:100 in 1× Reaction Lysis Buffer (Promega). 50 ml of diluted lysates were mixed with 50 μl of Firefly Luciferase Reagent, and luciferase activity was measured on a BMG Labtech Omega luminometer. To account for potential differences in transfection efficiency, luciferase values were normalized to β-galactosidase values. 50 μl of undiluted cell lysates were mixed with 50 μl of 2× Assay Buffer (Promega) and incubated for 30 minutes at 37°C. Reaction was terminated by adding 150 μl of 1 M sodium carbonate (Promega). β-galactosidase values were measured on a Tecan Spark microplate reader at 420 nm and normalized to a standard curve generated with the β-galactosidase provided by Promega.
Reaction lysis buffer
Manufactured by Promega
Reaction Lysis Buffer is a buffer solution used in molecular biology applications to facilitate the lysis or disruption of cells, tissues, or other biological samples. It is designed to efficiently release the contents of these samples, including nucleic acids, proteins, and other biomolecules, for subsequent analysis or purification.
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Lab products found in correlation
2 protocols using reaction lysis buffer
1
Measuring Transfected Cell Luciferase Activity
2
Measuring Transfected Cell Luciferase Activity
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Transfected 293T cells seeded in 12-well plates were harvested using 350 ml of Reaction Lysis Buffer from the Luciferase Assay System (Promega) per well. Cell lysates were diluted 1:100 in 1× Reaction Lysis Buffer (Promega). 50 ml of diluted lysates were mixed with 50 μl of Firefly Luciferase Reagent, and luciferase activity was measured on a BMG Labtech Omega luminometer. To account for potential differences in transfection efficiency, luciferase values were normalized to β-galactosidase values. 50 μl of undiluted cell lysates were mixed with 50 μl of 2× Assay Buffer (Promega) and incubated for 30 minutes at 37°C. Reaction was terminated by adding 150 μl of 1 M sodium carbonate (Promega). β-galactosidase values were measured on a Tecan Spark microplate reader at 420 nm and normalized to a standard curve generated with the β-galactosidase provided by Promega.
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