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ProFluor® Ser/Thr PPase Assay

Fluorescent, “Add-Mix-Read” Ser/Thr Phosphatase Assay

  • Works with protein phosphatase 1 (PP1), PP2A, PP2B and PP2C
  • Ideal for high-throughput phosphatase assays in 96- or 384-well plate formats
  • Minimal test compound interference

Size

Catalog number selected: V1260

$ 895.00 Your price: Log In

ProFluor® Ser/Thr PPase Assay
4 plate
$ 895.00
Your price: Log In
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Convenient Detection of Serine/Threonine Phosphatase Activity

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 produces excellent Z´ values (>0.8) 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 the provided phosphorylated bisamide rhodamine 110 peptide substrate (S/T PPase R110 Substrate) and Control AMC Substrate (control for compounds that may inhibit the protease reaction). 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

Specifications

You are viewing: V1260 Change Configuration

What's in the box?

Item Part # Size

Stabilizer Reagent

V306A 1 × 12μl

Protease Reagent

V309A 1 × 240μl

S/T PPase R110 Substrate

V324A 1 × 12μl

Control AMC Substrate

V325A 1 × 12μl

MgCl2, 1M

V330A 1 × 1ml

MnCl2, 20mM

V331A 1 × 1ml

CaCl2, 100mM

V332A 1 × 1ml

NiCl2, 100mM

V333A 1 × 1ml

5X Reaction Buffer B

V334A 1 × 6ml

5X Termination Buffer B

V335A 1 × 5ml

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

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Specifications

You are viewing: V1261 Change Configuration

What's in the box?

Item Part # Size

Stabilizer Reagent

V306B 1 × 24μl

Protease Reagent

V309B 1 × 480μl

S/T PPase R110 Substrate

V324B 1 × 24μl

Control AMC Substrate

V325B 1 × 24μl

MgCl2, 1M

V330A 1 × 1ml

MnCl2, 20mM

V331A 1 × 1ml

CaCl2, 100mM

V332A 1 × 1ml

NiCl2, 100mM

V333A 1 × 1ml

5X Reaction Buffer B

V334B 1 × 12ml

5X Termination Buffer B

V335B 1 × 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

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