Using FRAP to Quantify Changes in Transcription Factor Dynamics After Cell Stimulation: Cell Culture, FRAP, Data Analysis, and Visualization

Mapping SOX9 transcriptional dynamics during multi-lineage differentiation of human mesenchymal stem cells

“…We refer our readers to Govindaraj et al for the detailed explanation of TF-FRAP method and interpretation of data [13,16]. In short: A higher immobile fraction (IF) and/or a longer recovery halftime of A2 (residence time) are the indicators of increased SOX9 transcriptional activity.…”

Fluorescence Recovery after Photobleaching in Colloidal Science: Introduction and Application

“…127 The guar oligomer is retained by the hydrogel until it reaches the colonic environment when it is degraded by bacteria-secreted enzymes. The diffusion of guar oligomer inside the hydrogel was quantified by FRAP, which revealed that diffusion was greatly reduced compared with noninteracting 99 x Sustr et al 86,100 x Pinte et al 101 x Bruggen et al 102,103 x Nauman et al 104 x Prakash et al 105 x Balijepalli et al 106 x Jin-lei et al 107 x Grosskopf et al 16 x Govindaraj and Pos 4 x Hager et al 108 x Saito et al 109,110 x Sancataldo and Vetri 111 x Kvist et al 112 x Kindt et al 97 x Bui et al 113 x Kingsley al. 114 x Dey et al 115 x Devkota et al 116 x Waharte et al 117 x Sato et al 118 x Hashimoto et al 119 x probes and stayed constant over a couple of hours, resulting in a slow release.…”

“…Diffusion characteristics of diverse materials are critical for nonbiological as well as biological applications, such as drug delivery, cell culture, and tissue engineering. − The behavior of material components determines material stability and mechanical characteristics; similarly, diffusion of active chemicals in the bulk material determines their presentation or release. In 1827, Robert Brown observed the free migration of plant organelles expelled from pollen grains into a surrounding aqueous solution.…”

Temporal Dynamics and Stoichiometry in Notch Signaling - from Notch Synaptic Complex Formation to NICD Nuclear Entry

“…FRAP analysis. FRAP analysis was conducted as described (Govindaraj & Post, 2021) using Fiji (Schindelin et al, 2012) on the fluorescent signal within the photobleached ROI of the Notch synapse or plasma membrane after correcting the fluorescent signals for the inherent photobleaching due to imaging; the fluorescence intensity of the first 10 time points prior to FRAP were averaged and normalized to 1. The FRAP recovery curve was fitted using a single decay exponential from which the diffusion coefficient was estimated as D = (0.224 x r 2 )/ t1/2, where r is the radius of the bleached ROI and t1/2 the half-life of recovery (Kang et al, 2012).…”