Chemistry Reference
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Figure 7.1 Application of FCS to membrane
receptor mobility and ligand binding on living
cells (Adapted from [137]). (A) The detection
volume is positioned on the cell surface for
monitoring the binding of fluorescent ligands to
membrane receptors. (B) Fluorescence time
course for tetrametylrhodamine-labeled
C-peptide bound to membrane receptors on
cultured human renal tubular cells. (C)
Autocorrelation function of the fluorescence
intensity fluctuations shown in B. The observed
autocorrelation was fitted by the diffusion model
described by Equation 7.3. Two components of
receptor-bound C-peptide were observed.
Diffusion times and corresponding fractions
were t b1 ¼80 ms, y 1 ¼0.75; t b2 ¼1ms,
y 2 ¼
0.15. A small fraction of unbound C-peptide
that diffuses into the volume element extending
into the extracellular medium was characterized
by
0.10. (D) Distribution of
diffusion times calculated from the auto-
correlation function in C. (E) Percent of bound
ligand versus the total Rh-CP concentration.
Inset: Scatchard plot according to Equation 7.4.
t f ¼
0.15ms and y 3 ¼
7.4
Investigating Kinetics and Thermodynamics of Ligand
Receptor Interactions by FCS
-
7.4.1
Principles
The most common methods to evaluate receptor - ligand characteristics are the
radioligand binding assays, which require the use of
radioactively-labeled
 
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