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For PrPSc labelling in the double immunolabelling protocol, the EnVision protocol was preferred to the longer TSA method.
A single immunohistochemical protocol for PGP9.5 labelling of nerves was tested on spleen sections before incorporating the procedure in the double immunolabelling protocol.
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The PrPC immunolabelling protocol revealed a strong, granular, cytoplasmatic immunolabelling of Purkinje cells (Figure 2D) and neuronal immunolabelling in the granule cell layer of the cerebellum.
Double immunolabelling shows that aortic MCP-1 (red), TGF-β1 (green) immunolabeling, and their co-localization (merged image: yellow color) increase in the aged rat aortic wall, predominantly within the thickened intima in rats (Figure 1A, bottom right panel).
As a positive control for the selected ISH protocol and to allow comparison with PrPC immunohistochemistry, the ISH protocol and the PrPC immunolabelling protocol were performed on tissue sections from the cerebellum of a normal one month old lamb.
The remaining sections were processed for fluorescent double immunolabelling.
To detect lymphatic vessels, double immunolabelling with CD31 and the lymphatic marker Pdn was performed.
Double immunolabelling was thereafter performed using antibodies directed against HSPB1 and HLA-DR.
For 14 patients, there were no data on vimentin neoplastic cell expression because of tissue loss during the immunolabelling protocol.
Double immunolabelling confirmed this observation; C1q and C3b co-localized with GFAP + reactive astrocytes within the plaque and peri-plaque (C3b in this manuscript refers to immunolabelling with the C3b/iC3b-specific monoclonal antibody C3/30).
To differentiate VAMP1 and VAMP2 expression between excitatory and inhibitory axon terminals we performed double immunolabelling for each VAMP isoform and GAD65 (for inhibitory) and VGLUT1 (for excitatory).
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