Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot3822

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Probing Protein Interactions Using GFP and FRET

Stage 3: FLIM-FRET Measurements

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

The first 100 words of the full text of this article appear below.


INTRODUCTION

This stage presents a basic plan for capturing a series of images by fluorescent lifetime imaging microscopy. The protocol specifically describes data acquisition for a particular variant of GFP (EGFP) or Oregon Green as a donor fluorophore, but it can be adapted for image acquisition of other chromophore systems as described in the table below. This protocol was provided by Alisa G. Harpur and Philippe I.H. Bastiaens (Imperial Cancer Research Fund, London).


MATERIALS

Cells (live or fixed) expressing the probe proteins, prepared as described in Probing Protein Interactions Using GFP and FRET Stage 2: Cell Preparation for FLIM-FRET Analysis

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METHOD

1. Select . . . [Full Text of this Article]


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