| Laboratory Equipment
PerkinElmer, Inc.
Automated Liquid Handling
940 Winter St.
Waltham, MA 02451
(800) 762-4000 (US & Canada) or (+1) 203 925-4602
ctinfo@perkinelmer.com
www.perkinelmer.com/ATPLite_protocol
Automated Cytotoxicity and Cell Proliferation Assays
Using the JANUS Cellular Workstation with ATPlite 1step
ATPlite
TM
1step is a homogenous, luciferase-based luminescence
Typical Results:
assay system for the quantitative measurement of proliferation
For both compounds, the kinetics of the response was similar with
and cytotoxicity in cultured mammalian cells. We describe here
a greater degree of toxicity observed after 48 hours. Actinomycin D
a protocol for automating the ATPlite 1-step system using the
demonstrated approximately 10-fold higher potency compared to
JANUS
®
Cellular Workstation. ATPlite 1-step system is used here
Staurosporine. Dose-response curves were generated and Z´ values
to measure the cytotoxic dose-response of U937 cells, by exposing
determined. Sample data is shown in Figure 2 with an average Z´
cells to Staurosporine, a non-selective tyrosine kinase inhibitor, or
of 0.82 across 10 plates for U937 cells treated with Staurosporine.
Acitnomycin D, which binds DNA and blocks mRNA transcription by
For complete assay detail and full data sets, please download the
RNA polymerase. Results, presented in the full protocol, demonstrate
complete protocol.
the key performance and pharmacological parameters of the assay
with typical Z´ values between 0.82 and 0.85.
Hardware:
JANUS Cellular Workstation (Figure 1)
FlexDrop PLUS EX Precision Reagent Dispenser
Materials:
ATPlite 1step kit (PerkinElmer)
Cell lines
• HepG2 Cells
• U937 Cells
Growth medium [MEM (Invitrogen, Inc.), 10% FBS, 2 mM
L-Glutamine, 100 mM sodium pyruvate and 10 mM HEPES]
Figure 1
Staurosporine (Sigma, Inc.)
Actinomycin D (Sigma, Inc.)
384-well CulturPlate
TM
, white (PerkinElmer)
Protocol:
The general ATPLite 1step assay protocol is segmented into 5 steps:
Cell Seeding was conducted using the FlexDrop PLUS EX Precision
Reagent dispenser in stand-alone mode. All subsequent assay plate
processing was fully automated on the JANUS Cellular Workstation. A
general outline of each protocol step is provided in the full protocol.
1. Cell Seeding
2. Standards/Compound Addition
3. Assay Incubation
Figure 2
4. ATPlite 1step Reagent Addition
5. Assay Plate Reading
Protocol Guide ı 2008 View entire protocol online at www.biotechniques.com/protocol ı BioTechniques ı 41
Perkin Elmer Protocol.indd 41 10/26/07 11:42:09 AM
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