Rabbit anti mbp

Bielschowsky silver staining and immunohistochemistry were carried out according to standard procedures. The following primary antibodies were used: rabbit anti-MBP (1:1,000, Dako, Denmark), mouse anti-MBP peptide_70–89 (SMI94; 1:3,000, Covance, Princeton, New Jersey, USA), mouse anti-KiM1P (1:5,000) [43 (link)], rabbit anti-Olig2 (1:100, IBL, Spring Lake Park, Minnesota, USA), mouse anti-NogoA (1:10,000, mAb 11C7, a generous gift from M.E. Schwab, Brain Research Institute, ETH and University of Zurich, Switzerland) and mouse anti-Ki67 (1:50, clone Mib-1, Dako, Denmark). To identify Olig2⁺ or NogoA⁺ cells in control non-MS, normal-appearing MS and demyelinated MS neocortex, we performed double-immunolabeling combining either rabbit anti-Olig2 with mouse anti-MBP_70–89 (SMI94) or mouse anti-NogoA with rabbit anti-MBP.
Double-labeling immunohistochemistry was performed combining DAB (first primary antibody) and Fast Blue (second primary antibody with APAAP; Dako). For fluorescence double-labeling, Cy3- or Cy2-conjugated goat anti-mouse IgG and goat anti-rabbit IgG (both from Jackson ImmunoResearch Europe Ltd.) were used.

Cells were fixed with 4% paraformaldehyde. Cell-specific proteins were identified by indirect immunofluorescence using mouse anti-β-III-tubulin (1:2000; R&D Systems); rabbit anti-GFAP (1:500; Dako) and rabbit anti-MBP (1:250; Dako) antisera. The anti-rabbit DyLight 568- and anti-mouse AlexaFluor 488-labeled (1:500; Invitrogen) were used as secondary antisera. For nuclear staining, cells were incubated with 1 μg/mL Hoechst 33258.