Measurement of macromolecules interactions by AlphaScreen (PERKIN ELMER)

Introduction

AlphaScreen™ (Amplified Luminescent Proximity Homogeneous Assay), developed by PERKIN ELMER, is bead-based assay technology used to study biomolecular interactions in a microplate format.

This assay is homogeneous and requires only small amounts of protein.

Principle

Donor beads contain a photosensitizer, phthalocyanine, which converts ambient oxygen to an excited and reactive form of O2, singlet oxygen, upon illumination at 680 nm

If the donor bead is close to the acceptor bead (< 200 nm), energy is transferred from the singlet oxygen to thioxene derivatives within the Acceptor bead, subsequently culminating in light production at 520-620 nm

Beads can be coupled to many molecules allowing the capture of proteins fused to specific sequences.

AlphaScreen signal is proportional to the protein-protein interaction.

Protocole

Protein-protein interaction
Format384 wells
Protein 1 + Protein 2 +/- inhibitor
Reaction conditions to be​​​​​​​ defined  
Revelation

Donor/acceptor beads​​​​​​​ mix

Conditions to be defined

DetectionAlphaScreeen measurement

                                                                                                                                       

Notes :

  • Allow 50 µL at 10 mM or 3 mg of sample to test 5 concentrations (3 points/concentration)

Interaction of a purified protein with its receptor in the presence of an inhibitor

All models can be adapted to your request.