Team:Queens Canada/renillia

Renilla Luciferase Assay

Introduction

Luciferase isolated from Renilla reniformis, can be used as a genetic reporter to study receptor activity. Renilla luciferase, a monomeric 36kDa protein, catalyzes coelenterate-luciferin oxidation to produce light. Post-translational modification is not required for its activity, and the enzyme may function as a genetic reporter immediately following translation. The Renilla luciferase substrate, coelenterazine, also emits light from enzyme-independent oxidation, a process known as autoluminescence

Materials

• • 100 µl of Renilla Luciferase Assay Reagent
  • 20 µl of cell lysate

Procedure

• Manual Luminometer

1. Add 100 µl of Renilla Luciferase Assay Reagent to the luminometer tube.

2. Add 20 µl of cell lysate. Mix quickly by flicking the tube or vortexing for 1-2 seconds.

3. Place the tube in a luminometer and initiate measurement. Luminescence should be integrated over 10 seconds with a 2-second delay. Other integration times may be used. If the luminometer is not connected to a printer or computer, record the Renilla luciferase activity measurement.

4. Discard the reaction tube and proceed to the next sample, repeating Steps 1-3.