Team:Queens Canada/glo

Glo Lysis for Luciferase · Benchling

Glo Lysis for Luciferase

Introduction

Glo Lysis Buffer (GLB), 1X, is a proprietary formulation developed to promote rapid lysis (within 5 minutes) of cultured mammalian cells without scraping or performing freeze-thaw cycles. It is fully compatible with Bright-Glo™, Steady-Glo® ONE-Glo™ and Renilla-Glo™ Luciferase Assay Reagents for analysis of luciferase expression. The half-life of these reagents remains the same with or without the use of GLB; it is >5 hours for Steady-Glo® and >24 minutes for Bright-Glo™ Reagent.

Materials

  • Glo Lysis Buffer

Procedure

  • Protocol
  1. Equilibrate the Glo Lysis Buffer to room temperature before use
  1. Equilibrate the cells to room temperature. Aspirate the medium from the cells. Gently rinse with 1X PBS (cell rinse is optional).
  1. Add a sufficient volume of Glo Lysis Buffer to the sample well or plate to cover cells (see Table 1). When using lysate with other applications, account for the volume needed for that application.
  1. Rock the plate slowly several times to ensure complete coverage of the cells with Glo Lysis Buffer.
  1. Incubate for 5 minutes at room temperature to allow lysis to occur.
  1. Transfer the lysate to luminometer tubes, plate wells or vials and add Steady-Glo® or Bright-Glo™ Assay Reagent (Glo Lysis Buffer can be used in a 1:1 ratio with reconstituted Steady-Glo® or Bright-Glo™ Assay Reagent). Wait 5 minutes, then measure luminescence with a luminometer following the manufacturer’s instructions.
  • Note: Lysate prepared with Glo Lysis Buffer can be stored at –20°C or –70°C and is stable over several freeze-thaw cycles