2012 Promega catalogue

Life
Science
Catalog
2012
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Contents
Table of
Contents
Cell Signaling
70
ProFluor ® Ser/Thr PPase Assay
Product Size Cat.# Price ($)
ProFluor ® Ser/Thr PPase Assay 4 plate V1260 1196.00
For Research Use Only. Not for Use in Diagnostic Procedures.
8 plate V1261 1967.00
Description: The ProFluor ® Ser/Thr PPase Assay measures purified serine/
threonine protein phosphatase activity in a multiwell plate format and involves
“add-mix-read” steps only—ideal for high-throughput applications. The assay
works with protein phosphatase 1 (PP1), PP2A, PP2B and PP2C. The assay
begins with a standard phosphatase reaction performed with a provided
phosphorylated bisamide rhodamine 110 peptide substrate (S/T PPase R110
Substrate) and Control AMC Substrate that serves as a control for compounds
that may inhibit the protease reaction. Following the phosphatase reaction, a
termination buffer containing a protease reagent is added, which both stops the
phosphatase reaction and removes amino acids specifically from the nonphosphorylated
substrate, liberating highly fluorescent rhodamine 110. However,
phosphorylated substrate, however, is resistant to digestion by the protease
reagent and remains nonfluorescent. Thus, fluorescence intensity measured in
this assay is directly correlated with phosphatase activity. The assay produces
excellent Z´ values (>0.8) in either 96- or 384-well plate formats and easily
distinguishes known phosphatase inhibitors from other compounds.
Features:
• Achieve Highly Predictive Results: Robust Z´ values greater than 0.7 in
either 96- or 384-well plate formats.
• Minimal Test Compound Interference: Fluorescent signal from Rhodamine
110 is much greater than signals from by test compounds.
• Control Peptide Included: Use AAF-AMC control peptide to monitor
protease activity and reduce false-positive hits.
• Simplify Your Assays: Add-mix-read format reduces the 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 S/T PPase
R110 Substrate and Control AMC Substrate from light.
Protocol Part#
ProFluor ® Ser/Thr PPase Assay Technical Bulletin TB324
RFU
P P
No Phosphatase
P P
Protease Reagent
P P
Nonfluorescent
160,000
140,000
120,000
100,000
80,000
60,000
40,000
20,000
r 2 = 0.99
P P
S/T Phosphatase
Protease Reagent
Fluorescent
0
0 10 20 30 40 50 60 70 80 90100 Percent Nonphosphorylated Peptide
Effect of phosphopeptide content on fluorescence intensity. The
graph shows the average RFU obtained after a 90-minute digestion
of mixtures of nonphosphorylated S/T PPase R110 Substrate and
phosphorylated substrate as indicated to mimic a phosphatase titration.
4166MB
ProFluor ® Tyrosine Phosphatase Assay
Product Size Cat.# Price ($)
ProFluor ® Tyrosine Phosphatase Assay 4 plate V1280 1185.00
For Research Use Only. Not for Use in Diagnostic Procedures.
8 plate V1281 2040.00
Description: The ProFluor ® Tyrosine Phosphatase Assay measures purified
tyrosine phosphatase enzyme activity in a multiwell plate format and involves
“add-mix-read” steps only—ideal for high-throughput applications. Tyrosine
phosphatases tested with the assay include PTP-1B, CD45, LAR PTPase and
YOP-51. The assay begins with a standard phosphatase reaction performed
with a provided phosphorylated bisamide rhodamine 110 peptide substrate
(PTPase R110 Substrate) and Control AMC Substrate that serves as a control
for compounds that may inhibit the protease. Following the phosphatase
reaction, a termination buffer containing a protease reagent is added, which
simultaneously stops the phosphatase reaction and removes amino acids
specifically from the nonphosphorylated substrate, liberating highly fluorescent
rhodamine 110. Phosphorylated substrate, however, is resistant to digestion by
the protease reagent and remains nonfluorescent. Thus, fluorescence intensity
measured in this assay is directly correlated with phosphatase activity. The assay
produces excellent Z´ values (>0.7) in either 96- or 384-well plate formats
and easily distinguishes known phosphatase inhibitors from other compounds.
Features:
• Achieve Highly Predictive Results: Robust Z´ values greater than 0.8 in
either 96- or 384-well plate formats.
• Observe Minimal Test Compound Interference: Substrate used at
micromolar concentration. Rhodamine 110 fluorescent signal produced is
much greater than the fluorescent signal given off by test compounds.
• Control Peptide Included: Control peptide (AAF-AMC) included that is
used to monitor protease activity. Reduces false positive hits.
• Simplify Your Assays: Simple add-mix-read format reduces the number
of plate-handling steps to fewer than that required for other phosphatase
assays.
• Save Time: Minimal throughput time compared to the multiple steps and
lengthy incubations with other phoshatase assays.
• 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 PTPase
R110 Substrate and Control AMC Substrate from light.
Protocol Part#
ProFluor ® Tyrosine Phosphatase Assay Technical Bulletin TB334
For complete and up-to-date product information visit: www.promega.com/catalog