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The flow cytometry data was analysed with a one site competition model equation using Prism v4.01 (GraphPad Software, San Diego, CA, USA).
The data best fit a one site binding saturation curve with an average KD of 665±183 nM (four independent experiments, range 420 860 nM).
Km and kcat for 13-HPOT were calculated by fitting the data sets to a one site saturation model for simple ligand binding using SigmaPlot 8 (Sigma-Aldrich).
A one site total binding fit using Prism 5.04 for Windows was used to fit the curves and determine the dissociation constants.
The binding isotherms were fitted using nonlinear regression with a one site binding module available in GraphPad Prism version 4.00 for Windows (GraphPad Software, San Diego, CA).
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For the competitive binding assay, a one site-fit logIC50 curve was used.
Dissociation constants (KD) were determined by non-linear regression to a one site-specific binding model.
Discrimination between a one-site and a two-site ping-pong mechanism was based on considerable evidence.
Data were fit with a one-site binding model using Origin 7.0.
The binding data were subjected to nonlinear regression analysis using a one-site equilibrium binding equation with Prism software.
The data were analyzed using a one-site total binding regression algorithm of GraphPadPrism (GraphPad Software, San Diego, CA, USA).
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