Figure 1.

Characterisation of SP235 using transfected cells. (A) Western blotting analysis of the specificity of the anti-VPAC2 antibody SP235. Extracts from HEK 293 cells stably transfected to express either the human VPAC1 receptor (hVPAC1) or the human VPAC2 receptor (hVPAC2) were separated on 7.5% SDS–polyacrylamide gels and blotted onto PVDF membranes. Membranes were then incubated with the rabbit monoclonal anti-VPAC1 antibody (SP234) or with the rabbit monoclonal anti-VPAC2 antibody (SP235) at a dilution of 1:500. Blots were developed using ECL. Two additional experiments gave similar results. Ordinate, protein molecular weight marker (kDa). (B) Characterisation of SP235 by immunofluorescent staining of transfected cells. HEK 293 cells stably transfected to express either hVPAC1 or hVPAC2 were fixed and immunofluorescently stained with the anti-VPAC1 antibody (SP234) or the anti-VPAC2 antibody (SP235). Representative results from one of three independent experiments are shown. Scale bar, 20 μm.