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ProFluor® Src-Family Kinase Assay

Fluorescence-Based Detection of Src-Family Tyrosine Kinase Activity

  • Robust Z´ values greater than 0.7 in either 96- or 384-well plate formats
  • Add-mix-read format reduces the number of plate-handling steps
  • Non-Radioactive: No radioactive waste disposal and safety issues

Size

Catalog number selected: V1270

$ 895.00 Your price: Log in

ProFluor® Src-Family Kinase Assay
4 plate
$ 895.00
Your price: Log in

"Add-Mix-Measure" Convenience for High-Throughput Src-Family Kinase Screening

The ProFluor® Src-Family Kinase Assay measures the activity of purified Src-family tyrosine kinases (Src, Lck, Lyn, Fyn, and Hck tested) 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 Src-family kinase 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 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 inversely correlated with kinase activity. A control peptide (AAF-AMC) is included to control for compounds that may inhibit the protease and reduce false-positive hits.

Minimal Test Compound Interference

Rhodamine 110 fluorescent signal produced is much higher than the fluorescent signal given off by test compounds. The assay produces excellent Z´ values (>0.7) in either 96- or 384-well plate formats and easily distinguishes known Src-family kinase inhibitors from other compounds.

Specifications

What's in the box?

Item Part # SizeConcentrationAvailable Separately

rATP

P113C 1 × 250μl10mM View Product

Stabilizer Reagent

V306A 1 × 12μl

5X Reaction Buffer A

V307A 1 × 6ml

5X Termination Buffer A

V308A 1 × 5ml

Protease Reagent

V309A 2 × 240μl

Control AMC Substrate

V325A 1 × 12μl

MnCl2

V342A 1 × 1ml300mM

Sodium Vanadate

V343A 1 × 200μl100mM

Src-Family Kinase R110 Substrate

V344A 1 × 12μl

SDS

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Use Restrictions

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

Storage Conditions

BB

Store at –20°C.

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