2013 Promega catalogue

Life
Science
Catalog
2013
Worldwide Contact List
Cell Signaling
ProFluor ® Tyrosine Phosphatase Assay
Product Size Cat.# Price ($)
ProFluor ® Tyrosine Phosphatase Assay 4 plate V1280 1197
For Research Use Only. Not for Use in Diagnostic Procedures.
8 plate V1281 2061
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 higher 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
ProFluor ® Tyrosine Phosphatase Assay Technical Bulletin
Part#
TB334
Section
Contents
Table of
Contents
86
For complete and up-to-date product information visit: www.promega.com/catalog