Pharmacology EvaluationUsing instructions provided by the assay protocol for Kd determination, competition assays (Ki, IC50), and order of reagent, control, and compound addition to the plate, pharmacology data was generated to validate a fully automated assay procedure illustrated by Figures 5 and 6...
Thus, a careful choice of the design, a protocol optimization and a computerized analysis of the data can yield a dramatic improvement in the precision of the parameter estimation over more conventional approaches. 展开 关键词: Radioligand Assay Ligands Kinetics Computer Simulation Models, Biological...
To compare IP1 accumulation a homogeneous time-resolved fluorescence (HTRF) signalling assay was performed, adapting a previously described protocol41. The cells were washed 48 h after transfection with phosphate-buffered saline (PBS) and stimulation buffer (Cisbio) and subsequently incubated for 1...
The NanoBiT-based homogenous ligand-receptor binding assay is convenient for use and suitable for both the wild-type and mutant receptors, representing a novel tool for interaction mechanism studies of these receptors with their ligands. In the present chapter, we provide detailed protocols for ...
Ligand binding experiments between small chemicals and proteins and the evaluation of dissociation constants of their complexes in competitive binding assays often rely on displacement of reporter probes by the tested ligand. The most widely adopted protocol uses a fluorescent ligand which changes its emi...
LRR domain modification protocol with the MAB reagent The 2M urea was removed from the solubilized LRR using the Sephadex G-25 size-exclusion column chromatography. The resin was initially equilibrated with an equilibration buffer (21mM Tris-Cl pH 8.0, 1mM EDTA, 0.01% (v/v) NP-40, 5% (v/...
Using this protocol, FN-120 bound to the abembryonic surface of blastocysts (Fig. 2D), where mouse trophoblast differentiation initiates (Dickson, 1963; Kirby et al., 1967). Embryos that were cultured for 48 h and then similarly potentiated with FN-120 bound considerably less FN-120 (Fig. ...
See data sheet for detailed culturing and assay protocol. 背景(Background) Tumor necrosis factor ligand superfamily member 4 (TNFSF4) is also known as glycoprotein Gp34, OX40 ligand (OX40L), TAX transcriptionally-activated glycoprotein 1 and CD252, which belongs to the tumor necrosis factor fam...
It was found that a ratio in the range of 10:4 was suitable for measuring the heats evolved from EmrE binding to any of the substrates in any of the membrane mimetics. A modified solubilization protocol to achieve this concentration was employed as described above, after various parameters and...
Fluorescence polarization signals were collected using a Pherastar FSX, following the fluorescence anisotropy protocol (Ex 481 nm, Em 521), with measurements read for 0.5 s/cycle for a total of 200 flashes. Ligand binding was initiated by mixing receptor-enriched membranes (4 µg/mg ...