2012 Promega catalogue

Cell Signaling
Kinase-Glo ® Kinase-Glo Luminescent Kinase Assays
® Luminescent Kinase Assays
Product Size Cat.# Price ($)
Kinase-Glo ® Luminescent Kinase Assay 10 ml V6711 153.00
10 × 10 ml V6712 690.00
100 ml V6713 592.00
10 × 100 ml V6714 Pls. Enq.
Kinase-Glo ® Max Luminescent Kinase Assay 10 ml V6071 178.00
10 × 10 ml V6072 782.00
100 ml V6073 697.00
10 × 100 ml V6074 Pls. Enq.
Kinase-Glo ® Product Size Cat.# Price ($)
Kinase-Glo
Plus Luminescent Kinase Assay 10 ml V3771 168.00
10 × 10 ml V3772 753.00
100 ml V3773 653.00
10 × 100 ml V3774 Pls. Enq.
For Research Use Only. Not for Use in Diagnostic Procedures.
® Luminescent Kinase Assay 10 ml V6711 153.00
10 × 10 ml V6712 690.00
100 ml V6713 592.00
10 × 100 ml V6714 Pls. Enq.
Kinase-Glo ® Max Luminescent Kinase Assay 10 ml V6071 178.00
10 × 10 ml V6072 782.00
100 ml V6073 697.00
10 × 100 ml V6074 Pls. Enq.
Kinase-Glo ® Plus Luminescent Kinase Assay 10 ml V3771 168.00
10 × 10 ml V3772 753.00
100 ml V3773 653.00
10 × 100 ml V3774 Pls. Enq.
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: The Kinase-Glo ® Luminescent Kinase Assays are homogeneous
nonradioactive 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). The assay procedure involves
a single reagent addition 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 IC50 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 Km 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: Use with a wide range of kinases (including
lipid, sugar and alcohol kinases) and substrates (peptides, proteins, lipids,
sugars and alcohols).
• Obtain Reliable Results: Less interference from library compounds.
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 ® Description: The 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.
® Luminescent Kinase Assays are homogeneous
nonradioactive 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). The assay procedure involves
a single reagent addition 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 IC50 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 Km 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: Use with a wide range of kinases (including
lipid, sugar and alcohol kinases) and substrates (peptides, proteins, lipids,
sugars and alcohols).
• Obtain Reliable Results: Less interference from library compounds.
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 Part#
Kinase-Glo ® Protocol Part#
Kinase-Glo Luminescent Kinase Assay Platform Technical Bulletin TB372
® Luminescent Kinase Assay Platform Technical Bulletin TB372
A.
3.0 × 108 A.
3.0 × 108 For complete and up-to-date product information visit: www.promega.com/catalog
Luminescence (RLU) Luminescence (RLU)
2.5 × 108 2.5 × 108 2.0 × 108 2.0 × 108 1.5 × 108 1.5 × 108 1.0 × 108 1.0 × 108 0.5 × 108 0.5 × 108 0
0
r2 r =0.9993
2 =0.9993
20 40 60
ATP (µM)
80 100 120
B.
y = 280837x + 2 × 10
ATP (µM)
16
r2 2 × 10
= 0.9971
0
7
4 × 107 6 × 107 8 × 107 1 × 108 1.2 × 108 1.4 × 108 1.6 × 108 B.
y = 280837x + 2 × 10
0 100 200 300 400 500 600
ATP (µM)
16
r2 2 × 10
= 0.9971
0
7
4 × 107 6 × 107 8 × 107 1 × 108 1.2 × 108 1.4 × 108 1.6 × 108 0 100 200 300 400 500 600
Luminescent output correlates with amount of ATP. A direct
relationship exists between the luminescence measured with the
Kinase-Glo ® Assay systems and the amount of ATP in the reaction.
Panel A. Kinase-Glo ® Plus Assay. Panel B. Kinase-Glo ® Max Assay. The
Kinase-Glo ® Luminescent output correlates with amount of ATP. A direct
relationship exists between the luminescence measured with the
Kinase-Glo
Assay is linear to 10μM (data not shown).
® Assay systems and the amount of ATP in the reaction.
Panel A. Kinase-Glo ® Plus Assay. Panel B. Kinase-Glo ® Max Assay. The
Kinase-Glo ® Assay is linear to 10μM (data not shown).
ProFluor ® ProFluor PKA Assay
® PKA Assay
Product Size Cat.# Price ($)
ProFluor ® Product Size Cat.# Price ($)
ProFluor PKA Assay 4 plate V1240 1185.00
For Research Use Only. Not for Use in Diagnostic Procedures.
8 plate V1241 2040.00
® PKA Assay 4 plate V1240 1185.00
For Research Use Only. Not for Use in Diagnostic Procedures.
8 plate V1241 2040.00
Description: The ProFluor ® Description: The ProFluor PKA Assay measures protein kinase A activity
using purified kinase in a multiwell plate format and involves “add-mix-read”
steps only—ideal for high-throughput applications. The assay begins with a
standard kinase reaction performed with a provided PKA bisamide rhodamine
110 peptide substrate. Following the kinase reaction, a termination buffer
containing a protease reagent is added, which simultaneously stops the kinase
reaction and removes amino acids specifically from the nonphosphorylated
PKA substrate, liberating highly fluorescent rhodamine 110. Phosphorylated
PKA substrate, however, is resistant to digestion by the protease reagent and
remains nonfluorescent. Thus, fluorescence intensity measured in this assay is
inversely correlated with kinase activity. The assay produces excellent Z´-factor
values (>0.8) in either 96- or 384-well plate formats and easily distinguishes
known PKA inhibitors from other compounds.
Features:
• Achieve Highly Predictive Results: Robust Z´ values greater than 0.7 in
either 96- or 384-well plate formats.
• Observe Minimal Test Compound Interference: Fluorescent signal
from Rhodamine 110 is much stronger than fluorescent signals given off
by test compounds.
• Homogeneous: Add-mix-read format reduces the number of steps.
• Non-Radioactive: No radioactive waste disposal and safety issues.
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
Storage Conditions: Store the entire system at –20°C. Protect the PKA
R110 Substrate from light. For best results, make solutions fresh and use immediately.
System components should be thawed on ice and returned to –20°C
as soon as possible. The PKA R110 Substrate is provided in 100% DMSO and
therefore requires thawing at room temperature.
® PKA Assay measures protein kinase A activity
using purified kinase in a multiwell plate format and involves “add-mix-read”
steps only—ideal for high-throughput applications. The assay begins with a
standard kinase reaction performed with a provided PKA bisamide rhodamine
110 peptide substrate. Following the kinase reaction, a termination buffer
containing a protease reagent is added, which simultaneously stops the kinase
reaction and removes amino acids specifically from the nonphosphorylated
PKA substrate, liberating highly fluorescent rhodamine 110. Phosphorylated
PKA substrate, however, is resistant to digestion by the protease reagent and
remains nonfluorescent. Thus, fluorescence intensity measured in this assay is
inversely correlated with kinase activity. The assay produces excellent Z´-factor
values (>0.8) in either 96- or 384-well plate formats and easily distinguishes
known PKA inhibitors from other compounds. See figure on next page.
Features:
• Achieve Highly Predictive Results: Robust Z´ values greater than 0.7 in
either 96- or 384-well plate formats.
• Observe Minimal Test Compound Interference: Fluorescent signal
from Rhodamine 110 is much stronger than fluorescent signals given off
by test compounds.
• Homogeneous: Add-mix-read format reduces the number of steps.
• Non-Radioactive: No radioactive waste disposal and safety issues.
• Choose Your Configuration: Learn more about our custom options for
this product at: www.promega.com/myway/
Storage Conditions: Store the entire system at –20°C. Protect the PKA
R110 Substrate from light. For best results, make solutions fresh and use immediately.
System components should be thawed on ice and returned to –20°C
as soon as possible. The PKA R110 Substrate is provided in 100% DMSO and
therefore requires thawing at room temperature.
Protocol Part#
ProFluor ® Protocol Part#
ProFluor PKA Assay Technical Bulletin TB315
® PKA Assay Technical Bulletin TB315
6979MB
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