2013 Promega catalogue

Life
Science
Catalog
2013
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Cell Signaling
Kinase-Glo ® Luminescent Kinase Assays
Product Size Cat.# Price ($)
Kinase-Glo ® Luminescent Kinase Assay 10 ml V6711 155
10 × 10 ml V6712 697
100 ml V6713 598
10 × 100 ml V6714 Pls. Enq.
Kinase-Glo ® Max Luminescent Kinase Assay 10 ml V6071 180
10 × 10 ml V6072 790
100 ml V6073 704
10 × 100 ml V6074 Pls. Enq.
Kinase-Glo ® Plus Luminescent Kinase Assay 10 ml V3771 170
For Research Use Only. Not for Use in Diagnostic Procedures.
10 × 10 ml V3772 761
100 ml V3773 660
10 × 100 ml V3774 Pls. Enq.
Description: The Kinase-Glo ® Luminescent Kinase Assays are homogeneous
non-radioactive methods for determining the activity of purified kinases by
quantifying the amount of ATP remaining in solution following a kinase reaction.
The assays are designed for use with multiwell plate formats, making them
ideal for automated high-throughput screening (HTS), and they can be used to
assay protein, lipid and sugar kinases. The assay procedure involves addition of
a single reagent directly to a completed kinase reaction. This addition results in
the generation of a luminescent signal correlated with the amount of ATP present
and inversely proportional to the amount of kinase activity. The Kinase-Glo ®
Assays generate a “glow-type” luminescent signal produced using a patented
stabilized luciferase (Ultra-Glo Luciferase) coupled with a proprietary buffer
system. When assayed in the presence of kinase reaction buffers, such as the
reaction buffer for PKA, the half-life of the luminescent output is greater than
five hours, eliminating the need for luminometers with injectors and allowing
for batch plate processing. The assay produces excellent Z´-factor values of
greater than 0.7 in 96- and 384-well formats, easily detects known kinase inhibitors
and provides IC 50 values comparable to those reported in the literature.
The Kinase-Glo ® Assay systems are differentiated by their linear response
to ATP (see figure below). The original Kinase-Glo ® Assay is linear to 10μM
ATP, while Kinase-Glo ® Plus Assay is linear to 100μM ATP. The newest assay,
Kinase-Glo ® Max, is linear to 500μM ATP, making it well suited for use with
kinases with high K m for ATP as well as for screening for kinase inhibitors that
do not compete at the ATP binding site.
Features:
• Assay a Variety of Kinases: Can be used for a wide range of kinases (including
lipid, sugar and alcohol kinases) and substrates (peptides, proteins,
lipids, sugars and alcohols).
• Obtain Reliable Results: Luminescence is much less susceptible to
interference from library compounds than other luciferase-based ATP detection
reagents. Z´-factor values greater than 0.7 in either 96- or 384-well
plate formats.
• Simplify Your Assay: Homogeneous—everything is performed in a
single well.
• Non-Radioactive: No radioactive waste disposal and safety issues.
• Automate This Assay: Validated automated methods available at: www.
promega.com/automethods/
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
• Screen for Non-ATP Binding Site Inhibitors: Use ATP concentrations
as high as 500μM (Kinase-Glo ® Max Assay).
Storage Conditions: Store at –20°C. Avoid multiple freeze-thaw cycles and
exposure to frequent temperature changes. These fluctuations can greatly alter
product stability.
Protocol
Kinase-Glo ® Luminescent Kinase Assay Platform Technical Bulletin
A.
Luminescence (RLU) Luminescence (RLU)
B.
20 40 60 80 100 120
2.5 × 10 8 r 2 =0.9993
2.0 × 10 8
1.5 × 10 8
1.0 × 10 8
0.5 × 10 8
0
0
3.0 × 10 8 ATP (µM)
ATP (µM)
Part#
TB372
1.6 × 10 8
1.4 × 10 8 y = 280837x + 2 × 10 16
1.2 × 10 8 r 2 = 0.9971
1 × 10 8
8 × 10 7
6 × 10 7
4 × 10 7
2 × 10 7
0
0 100 200 300 400 500 600
Luminescent output correlates with amount of ATP. A direct
relationship exists between luminescence measured with the Kinase-Glo ®
Assay systems and the amount of ATP in the reaction. Panel A. Data from
Kinase-Glo ® Plus Assay. Panel B. Data from Kinase-Glo ® Max Assay. The
Kinase-Glo ® Assay is linear to 10µM (data not shown).
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