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ProFluor® Tyrosine Phosphatase Assay

Fluorescent, “Add-Mix-Read” Tyrosine Phosphatase Assay

  • Tyrosine phosphatases tested include PTP-1B, CD45, LAR PTPase and YOP-51
  • Ideal for high-throughput applications in 96- or 384-well plate formats
  • Minimal test compound interference

Size

Catalog number selected: V1280

$ 895.00 Your price: Log In

ProFluor® Tyrosine Phosphatase Assay
4 plate
$ 895.00
Your price: Log In
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High-Throughput Detection of Tyrosine Phosphatase Activity

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 include PTP-1B, CD45, LAR PTPase and YOP-51. 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.

Protocol Overview

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 (control for compounds that may inhibit the protease). Following the phosphatase reaction, a termination buffer containing a protease reagent is added, which stops the phosphatase reaction and removes amino acids specifically from the nonphosphorylated substrate, liberating highly fluorescent rhodamine 110. Phosphorylated substrate is resistant to digestion by the protease reagent and remains nonfluorescent. Thus, fluorescence intensity measured in this assay is directly correlated with phosphatase activity.

Advantages

  • Minimal Test Compound Interference. The Rhodamine 110 fluorescent signal produced is much higher than the fluorescent signal given off by test compounds.
  • Control Peptide Included (AAF-AMC) for monitoring protease activity, reducing false-positive hits.
  • Simple add-mix-read format reduces the number of plate-handling steps to fewer than that required for other phosphatase assays.
  • No radioactive waste disposal and safety issues.
  • Custom formats available: www.promega.com/custom

Protocols

Specifications

You are viewing: 4 plate Change Configuration

What's in the box?

Item Part # SizeConcentration

Stabilizer Reagent

V306C 1 x 60μl

Protease Reagent

V309A 1 x 240μl

Control AMC Substrate

V325A 1 x 12μl

5X Reaction Buffer B

V334A 1 x 6ml

Sodium Vanadate

V343A 1 x 200μl100mM

PTPase R110 Substrate

V345A 1 x 12μl

5X Termination Buffer C

V346A 1 x 5ml

SDS

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

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Specifications

You are viewing: 8 plate Change Configuration

What's in the box?

Item Part # SizeConcentration

Stabilizer Reagent

V306D 1 x 120μl

Protease Reagent

V309B 1 x 480μl

Control AMC Substrate

V325B 1 x 24μl

5X Reaction Buffer B

V334B 1 x 12ml

Sodium Vanadate

V343A 1 x 200μl100mM

PTPase R110 Substrate

V345B 1 x 24μl

5X Termination Buffer C

V346B 1 x 10ml

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

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