We compared three radioligands for use in a cytoplasmic estrogen-receptor assay, using pooled cytosol from human breast adenocarcinomas. The estrogen receptor content was determined in vitro by a dextran-coated charcoal method involving a 4-h incubation with and without diethylstilbestrol. Tritiated moxestrol failed to come to equilibrium in 4 h, thereby preventing the use of conventional one-c...

This work evaluates the use of mobility ratios (M) to estimate binding constants of proteins to ligands using affinity capillary electrophoresis (ACE). This concept is demonstrated using two model systems: vancomycin (Van) from Streptomyces orientalis and carbonic anhydrase B (CAB, EC 4.2.1.1). A plot of change in M (deltaM) over the concentration of ligand [L] versus deltaM yields a more usefu...

In the present work, reusable magnetic molecularly imprinted polymers have been used for first time as an adsorbent determination of 2-phenylphenol. Magnetic selective separation and preconcentration 2-phenylphenol native fluorescence has sensitive analyte. The developed method is rapid was completed in 40 minutes. Limit detection (LOD) found to be 1.09 µg L-1 imprinting factor 1.87. linearity ...

This investigation focuses on the effectiveness of coconut shell activated carbon in removing synthetic dye mixture from aqueous solutions. Coconut carbon, an economical and effective adsorbent, was made agricultural waste raw material chemically by sulphuric acid treatment. Activated is characterised using FT-IR SEM analysis. Batch adsorption studies were conducted adjusting conditions such co...

In this two-point Scatchard-plot assay, with which a test of competitive inhibition is combined, the sample is mechanically homogenized in a buffer containing dithiothreitol, ultracentrifuged to obtain a fat-free cytosol, the protein content of which is then adjusted, and free and bound labeled estradiol are separated with dextran-coated charcoal after overnight incubation. We tested the method...

Previous studies of the kinetics of steroid binding to receptors have shown a lack of agreement between the dissociation constant obtained at equilibrium by Scatchard analysis (Scatchard, G. (1949) Ann. N. Y . Acad Sci. 51, 660-672) and that calculated using the rate constants of dissociation and association (kd/k,) (Pratt, W. B., Kaine, J. L., and Pratt, D. V. (1975) J. Biol. Chem. 250, 4584-4...

One of the best approaches to date to obtain overall binding constants (Ko) for Al and dissolved organic matter (DOM) from acidic soil solutions is to collect 'free' Al data with diffusive gradients in thin films (DGT) and to infer the Ko values by fitting a continuous distribution model based on Scatchard plots. Although there is clear established literature demonstrating the usefulness of the...

Two distinct thyrotropin (TSH) binding species have been separated from solubilized porcine thyroid membranes. Membranes was solubilized with 1% Triton X-100, and the supernatant was recovered by centrifugation at 105,000 X g. Scatchard analysis of thyrotropin binding to solubilized membranes (SM) yielded a nonlinear plot with Kd values for the high and low affinity components similar to those ...

High-affinity [3H]serotonin binding activity has been solubilized from bovine cerebral cortical membranes by using Triton X-100, Tween-80, and octyl-beta-D-glucopyranoside. This mixture of detergents solubilizes the high-affinity [3H]serotonin binding activity present in crude membrane preparations with retention of 75-90% specific binding. The detergent mixture was chosen because it can easily...