Luciferase activity in cell lysates was measured using the Luciferase Assay System (Promega). Cells were washed with PBS and harvested 72 h after transfection using 50 µl of lysis buffer. Cell extracts were centrifuged and 30 µl of the supernatant was mixed with 50 µl of luciferase assay buffer. Luciferase activity was measured with the GloMax-Multi+ Microplate Multimode Reader luminometer (Promega Corp). β-galactosidase activity was assayed using 1 mg of o-nitrophenyl β-D-galactopyranoside (AmResco) as the substrate in buffer Z (60 mM Na2HPO4, 40 mM NaH2PO4, 10 mM KCl, 1 mM MgSO4, 0.07% β-mercaptoethanol) and incubated at 37°C until yellow staining. The product was determined by absorption at 420 nm. This value was used to correct variations in transfection efficiency. Luciferase results were calculated as the ratio of luciferase activity per unit of β-galactosidase activity. Duplicate samples were analyzed for each data point.
Glomax multi microplate multimode reader luminometer
Manufactured by Promega
The GloMax-Multi+ Microplate Multimode Reader is a luminometer designed to measure luminescent signals in microplates. It is capable of detecting a wide range of luminescent reporters and can be used for a variety of applications that require luminescent detection.
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Lab products found in correlation
2 protocols using glomax multi microplate multimode reader luminometer
1
Luciferase Activity Assay in Cell Lysates
2
Luciferase Assay for Transfection Efficiency
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Luciferase Assay System (Promega) was used to measure luciferase (Luc) activity in cell lysates. Seventy-two hours after transfection, cells were washed with PBS and harvested using 50 μL of lysis buffer provided by the manufacturer. GloMaxMulti + Microplate Multimode Reader Luminometer (Promega) was used to measure luciferase activity. β-Galactosidase activity was assayed using 1 mg of o-nitrophenyl β-d-galactopyranoside (AmResco) as the substrate in buffer Z (40 mM NaH 2 PO 4 , 60 nM Ns 2 HPO 4 , M Schanton and others 594 Reproduction (2020) 160 591-602 10 nM KCl, 0.07% β-mercaptoethanol, 1 mM MgSO 4 ). Samples were incubated at 37°C until yellow staining. Enzymatic reaction was stopped with 75 μL of sodium carbonate. Color development was determined by absorption at 420 nm. The variation in transfection efficiency was corrected with this value. Luc results were calculated as the percentage of Luc activity per unit of β-galactosidase activity. For each data duplicates were performed.
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