RealTime-Glo™ MT Cell Viability Assay

Why Use RealTime-Glo MT Cell Viability Assay?

With the RealTime-Glo™ MT Cell Viability Assay, you can monitor cell viability continually in the same sample well out to 72 hours to obtain more information about the mode of action of a treatment with regard to time and dose dependence. The assay reagent can be added to cells during plating, during treatment or at the end of treatment. No cell washing, media removal or further additions are required.

The nonlytic nature and rapid response of the assay make it possible to monitor cell viability over time in the same well, and enables easy multiplexing with other cell-based assays or downstream applications. The assay measures the reducing potential of viable cells, and is ATP-independent, providing an orthogonal method for viability or cytotoxicity determination.

The RealTime-Glo Assay Measures Reducing Potential: A Well-Established Indicator of Cell Viability

Assay Principle

In the RealTime Glo MT Cell Viability Assay, a non-lytic NanoLuc Luciferase reaction occurs in the culture medium. NanoLuc Luciferase and MT Cell Viability Substrate are added to cell culture media. The MT Cell Viability Substrate then diffuses into cells where it is reduced to form a NanoLuc Substrate, which exits the cell and is used rapidly by NanoLuc Luciferase in the media. Only metabolically active cells can reduce the substrate, and light production is proportional to the number of live cells in culture. Dead cells cannot reduce the substrate.

realtime-glo detects live cells 12420MB-W

Save Time, Cell Culture and Reagent Costs

One RealTime-Glo Assay Plate Yields the Same Data as Many End-point Assay Plates

Five hundred A549 cells were plated in medium containing the RealTime-Glo MT Reagents in 384-well plates and various concentrations of bortezomib. The luminescence signal was determined from each well every hour for 72 hours in a Tecan M200 plate reader with gas control module (37 degrees C/5% CO2).

Real time live cell data collection 12466MF-W-a

bortezomib-live-dead-cell data-12466MF-W-b

Data from specific time points plotted to calculate EC50 at these time points.

Rapidly Detect Changes in Cell Viability

The RealTime-Glo MT Cell Viability Assay generates a luminescent signal that is dependent upon continuous reduction of the MT cell viability substrate by viable cells and rapid turnover by NanoLuc Luciferase. Conventional assays based on reducing potential (alamarBlue, MTT) are accumulation-based and do not allow real-time detection of cell death in the same well. In this experiment, A549 cells were treated with 1% Triton X-100. Viability assay reagents were applied and incubated for the indicated times post-treatment.

alamar blue Realtime-Glo comparison-12613MB-W

Add to Cells at Plating, Dosing, or Endpoint

Flexible Assay Setup

Perform real-time cell viability monitoring or endpoint detection. You decide how to set up your assay. Add the RealTime-Glo Reagent to cells at any time point when cell viability readings are desired. The assay is compatible with high-throughput screening in 96-, 384- and 1536-well plates.

Realtime kinetic or endpoint viability methods 12474MB-W

Multiplex with Other Assays for More Information Per Sample

The nonlytic, live-cell RealTime-Glo Assay is compatible with downstream applications and allows multiplexing with other assays, conserving precious samples and providing more informative data per sample.

Realtime-Glo viability data-etoposide-12573MD-W-a

Realtime-cytotoxicity-etoposide-data-12573MD-W-b

Protocols

Complete Protocol

RealTime-Glo™ MT Cell Viability Assay Technical ManualPDF (1809 KB) – English

What's in the box?

Item	Part #	Size
NanoLuc® Enzyme, 1,000X	E499A	1 × 10μl
MT Cell Viability Substrate, 1,000X	G971A	1 × 10μl

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Patents and Disclaimers

Patent Pending.

What's in the box?

Item	Part #	Size
NanoLuc® Enzyme, 1,000X	E499A	10 × 10μl
MT Cell Viability Substrate, 1,000X	G971A	10 × 10μl

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Patents and Disclaimers

Patent Pending.

What's in the box?

Item	Part #	Size
NanoLuc® Enzyme, 1,000X	E499B	1 × 100μl
MT Cell Viability Substrate, 1,000X	G971B	1 × 100μl

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Patents and Disclaimers

Patent Pending.

Resources

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RealTime-Glo™ MT Cell Viability Assay

Monitor Cell Viability in Real Time

Why Use RealTime-Glo MT Cell Viability Assay?