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Analysis of molecular binding in living cells with FCCS

Interactions are directly evident by the presence of a cross-correlation between two spectrally distinct channels. Furthermore, the ratio of free versus bound fraction in equilibrium reactions can be quantified. FCCS measurements are conducted with the FCS wizard.

Example
Caspase6, a downstream effector of the TNF-dependent apoptosis pathway, dimerizes upon activation (shown in the graph). A GPF-labeled construct and an mCherry-labeled construct were expressed in HeLa cells. 50% of the resulting double-labeled dimers (grey box) are detected in the cross-correlation signal. FCCS thus proves the active form of Caspase.

We thank Prof. Dr. Roland Eils and Dr. Joel Beaudouin, Bioquant, University of Heidelberg, Germany.

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