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. Author manuscript; available in PMC: 2015 Mar 20.
Published in final edited form as: Chem Biol. 2014 Feb 27;21(3):338–344. doi: 10.1016/j.chembiol.2014.01.009

Figure 2. In vitro CB2R saturation binding assay of IR700DX-mbc94.

Figure 2

Cells were preincubated with (non-specific binding) or without (total binding) 1 μM of the blocking agent SR144528 for 1 h, and then incubated overnight with an increasing concentration of IR700DX-mbc94 at 37 °C. Cells were then rinsed and the fluorescence intensity was recorded. The specific binding was obtained by the subtraction of non-specific binding from the total binding. The dissociation constant (Kd) and receptor density (Bmax) were estimated from the non-linear fitting of specific binding versus IR700DX-mbc94 concentration using Prism software. Each data point represents the mean ± SD based on triplicate samples.