Hi everyone,

I have a few questions about using TZM-bl cells with a high RLU background.

My Protocol:

• Day 1: Plate TZM-bl cells in a 96-well (half-area) plate.
• Day 2: Lyse the cells with lysis buffer for 10 minutes with shaking.
• Then, transfer 25 µL of cell lysate to a white 96-well (half-area) plate and add 25 µL of Bright-Glo assay reagent (substrate + assay buffer).
• Measure the RLU.

The Problem:

The RLU of my uninfected cells is approximately 10^4, which is very high. This control is typically below 1000. We initially suspected contamination, but a new vial of cells and a newly purchased vial of Bright-Glo both yielded the same high background signal.

Has anyone else encountered this issue? What were your solutions?

Thanks in advance for your help!