Assay Method Information

Assay Name:  Receptor Binding Assay
Description:  In general, the results are expressed as a percent of control specific binding: measured⁢specific⁢bindingcontrol⁢specific⁢bindingĂ—100and as a percent inhibition of control specific binding:100-(measured⁢specific⁢bindingcontrol⁢specific⁢bindingĂ— 100)obtained in the presence of the test compounds. The IC50 values (concentration causing a half-maximal inhibition of control specific binding) and Hill coefficients (nH) are determined by non-linear regression analysis of the competition curves generated with mean replicate values using Hill equation curve fitting:Y=D+[A-D1+(C/C5⁢0)nH] where Y=specific binding, A=left asymptote of the curve, D=right asymptote of the curve, C=compound concentration, C50=IC50, and nH=slope factor. This analysis was performed using in-house software and validated by comparison with data generated by the commercial software SigmaPlot 4.0 for Windows ( 1997 by SPSS Inc.). The inhibition constants (Ki) were calculated using the Cheng Prusoff equation: Ki=IC5⁢0(1+L/KD) where L=concentration of radioligand in the assay, and KD=affinity of the radioligand for the receptor.
Affinity data for this assay
 

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