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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: J Neurochem. 2011 Nov 24;120(1):56–69. doi: 10.1111/j.1471-4159.2011.07559.x

Fig. 8. Detection of plasma membrane receptors expressed at the cell surface in the detergent-resistant membrane (DRM) fraction.

Fig. 8

HEK293T cells were transfected with cDNA for the N-terminal FLAG-tagged D2R or MOR or empty vector (control) and 48 hr post-transfections were incubated for 1 hr at 37 °C with an anti-FLAG antibody targeting the extracellular FLAG epitope on the respective receptors. The cells were subsequently washed three times in ice cold PBS, treated for 1 hr with 2% v/v (20mL/L) TX100 at 4 °C and the TX100-insoluble and soluble fractions were separated by centrifugation as outlined in the Materials and Methods section. A. The receptor-targeting antibody in TX100 detergent soluble (S) and insoluble (P, pellet) cell fractions visualized by immuno-blotting, using appropriate secondary antibodies, as described in the Materials and Methods sections. “control” refers to a control group of cells transfected with empty plasmid vector and then incubated with anti-FLAG antibody. B. Levels of extracellular epitope targeting FLAG antibody in TX100-soluble (closed black bars) and insoluble (open bar) cellular fractions visualized in A and expressed as percentage of the total antibody detected in both the TX100 soluble and insoluble fractions (mean±SEM, n=5, *p<0.02).