RealTime-Glo™ MT Cell Viability Assay

Live / Dead assay mammalian cells - THP-1

Experiment
Live / Dead assay mammalian cells - THP-1
Product
RealTime-Glo™ MT Cell Viability Assay from Promega
Manufacturer
Promega

Protocol tips

Protocol tips
Add an equal volume of 2X RealTime-Glo™ reagent to the cells.

Incubate the cells for 10–60 minutes in the cell culture incubator

Publication protocol

Commercial Availability
Commercial kits containing the pro-substrate and the engineered shrimp-derived luciferase are available from Promega Corporation. RealTime-Glo™ MT Cell Viability Assay, Cat.# G9711 (100 reactions); G9712 (10x100 reactions); G9713 (1000 reactions)

Reagent Preparation and Real Time Viability Assay Protocol
The MT Cell Viability Substrate and the NanoLuc® Enzyme are both supplied at 1000X the final recommended concentration.

For continuous read mode:

1.Equilibrate the MT Cell Viability Substrate and the NanoLuc® Enzyme to 37°C.

2.Harvest cells and adjust to desired cell density to be used in the assay.

3.Add MT Cell Viability Substrate and the NanoLuc® Enzyme to the cell suspension.

4.Dispense cell suspension containing MT Cell Viability Substrate and the NanoLuc® Enzyme into white opaque walled multiwell plates suitable for luminescence measurements.

5.Add test compound and incubate for desired length of time.

6.Record luminescence.

For endpoint mode:

1.Harvest cells and adjust to desired cell density to be used in the assay.

2.Add test compound to cells and incubate for desired length of time.

3.Equilibrate the MT Cell Viability Substrate and the NanoLuc® Enzyme to 37°C.

4.Dilute MT Cell Viability Substrate and the NanoLuc® Enzyme in cell culture medium to form 2X RealTime-Glo™ Reagent.

5.Add an equal volume of 2X RealTime-Glo™ Reagent to cells.

6.Incubate at 37°C for 10-60 min.

7.Record luminescence.

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Papers

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Manufacturer protocol

Download the product protocol from Promega for RealTime-Glo™ MT Cell Viability Assay below.

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