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    ARTICLE

    An Improved Mathematical Approach for Determination of Molecular Kinetics in Living Cells with FRAP

    Tanmay Lele1,1, Philmo Oh1,1, Jeffrey A. Nickerson1,1,2,2, Donald E. Ingber1,1,3,3

    Molecular & Cellular Biomechanics, Vol.1, No.3, pp. 181-190, 2004, DOI:10.3970/mcb.2004.001.181

    Abstract The estimation of binding constants and diffusion coefficients of molecules that associate with insoluble molecular scaffolds inside living cells and nuclei has been facilitated by the use of Fluorescence Recovery after Photobleaching (FRAP) in conjunction with mathematical modeling. A critical feature unique to FRAP experiments that has been overlooked by past mathematical treatments is the existence of an `equilibrium constraint': local dynamic equilibrium is not disturbed because photobleaching does not functionally destroy molecules, and hence binding-unbinding proceeds at equilibrium rates. Here we describe an improved mathematical formulation under the equilibrium constraint which provides a more… More >

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