Intact cell binding

Saponin solution (140 mM potassium glutamate-HCl, pH 6.8, with 1 mg/mL ATP and 0.1 mg/mL saponin) for the intact cell-binding assay. 

Using any of the carrier proteins available in highly purified form, eg, TBG or TBPA, a convenient and accurate quantitative determination of and is possible by displacement of radioiodinated or T. This procedure enables their quick determination at low concentrations even in the presence of coundess other substances that occur in body duids (31). In a similar fashion, intact cell nuclei or solubilized proteins from rat fiver cell nuclei, which display high affinities for thyroid hormones, especially T, have been used to establish relative binding affinities of many thyromimetic compounds (7). 

Analysis of Ligand Binding in the Intact Cell. A model of LR dynamics in the intact cell consistent with the results of these studies is presented in Figure 6, and a fit to the data of a polarization binding experiment is shown. The model has these characteristics ... 

Tsomides, T.J., Walker, B.D., and Eisen, H.N. (1991) An optimal viral peptide recognized by CD8+T cells binds very tightly to the restricting class I major histocompatibility complex protein on intact cells but not to the purified class I protein. Proc. Natl. Acad. Sci. USA 88, 11276-11280. 

Stein RA, Wilkinson JC, Guyer CA, Staros JV. 2001. An analytical approach to the measurement of equilibrium binding constants application to EGF binding to EGF receptors in intact cells measured by flow cytometry. Biochemistry 40 6142-6154. 

Bylund, D.B., Deupree, J. D., Toews, M.L. Radioligand-binding methods for membrane preparations and intact cells, in Methods in Molecular Biology, vol. 259 Receptor Signal Transduction Protocols, ed. Wdlars,... 

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