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 accurate estimate of molecular reaction… More >
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