Cells were washed with 1× PBS and then lysed in Luciferase Lysis Buffer (20 mM Tris pH 7.4 and 0.1% Triton X-100). Lysate was briefly vortexed and centrifuged at maximum speed for 15 min at 4°C. Enzymatic activity was measured in each sample on a luminometer using Luciferase assay substrate (E1501, Promega) or Renilla assay substrate (E2820, Promega). Immunoblots of luciferase extracts contained equal volumes from each triplicate.
Renilla assay substrate
Manufactured by Promega
The Renilla assay substrate is a reagent used in bioluminescent assays to measure the activity of the Renilla luciferase reporter gene. It provides the necessary substrate for the Renilla luciferase enzyme to catalyze a light-emitting reaction, which can be quantified to determine the expression level of the reporter gene.
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Lab products found in correlation
2 protocols using renilla assay substrate
1
Luciferase Activity Measurement Protocol
2
Luciferase Reporter Assay Protocol
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Transcriptional reporter assays were performed using luciferase reporter plasmids along with expression constructs (indicated in the figure legends) and a Renilla plasmid (pRL-Renilla, Promega) as an internal control. Cells were washed with 1× phosphate-buffered saline and harvested in Luciferase Lysis Buffer (20 mM Tris pH 7.4 and 0.1% Triton X-100). Enzymatic activity was measured in each sample on a luminometer (Lumat LB, Berthold) using Luciferase assay substrate (E1501; Promega) or Renilla assay substrate (E2820; Promega). Luciferase activity values obtained were normalized to Renilla activity in the same cell extracts and expressed as fold activation to the control.
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