Figure 2. Histoimmunoprecipitation and antigen identification

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Figure 2. Histoimmunoprecipitation and antigen identification Histoimmunoprecipitation and antigen identification Cryosections of rat or pig cerebellum were incubated with the patient's serum (1:100), washed in phosphate-buffered saline, and solubilized using 1% Triton X-100. The solution was incubated with protein-G-coated magnetic beads. The immunocomplexes were eluted by sodium dodecyl sulfate (SDS) and subjected to SDS–polyacrylamide gel electrophoresis (PAGE) analysis and Western blot. (A) SDS-PAGE and staining with colloidal Coomassie. Lane 1: molecular weight marker. Lane 2: histoimmunoprecipitate of the patient serum (cerebellum rat). Arrows indicate the position of the immunoprecipitated antigen at 113 kDa while dotted arrows indicate the position of immunoglobulin G heavy and light chain at 52 kDa and 27 kDa, respectively. Lanes 3, 4: rat cerebellum precipitated with sera exhibiting as yet undefined neuroimmune reactions (controls). (B) Western blot analysis with polyclonal rabbit anti-GluRδ2 antibody. Lane 1: molecular weight marker; lanes 2, 3: histoimmunoprecipitate of the patient's serum (cerebellum rat); lanes 4, 5: rat cerebellum precipitated with sera exhibiting as yet undefined neuroimmune reactions (controls); lanes 6, 7: histoimmunoprecipitate of the patient's serum (cerebellum monkey); lanes 8, 9: monkey cerebellum precipitated with sera exhibiting as yet undefined neuroimmune reactions (controls). Ramona Miske et al. Neurol Neuroimmunol Neuroinflamm 2016;3:e255 © 2016 American Academy of Neurology