Fixing and Permeabilization of cells: Absolute methanol (10 minutes in ice) and 0.1% Tween-20 in PBS, Blocking: 1% BSA, Primary antibody: Mouse Monoclonal antibody to MBP (cat # M-1384-50, 2 µg per ~10^6 cells) for 30 minutes at room temperature, Secondary antibody: Goat anti-mouse PE labeled secondary antibody (1:100 fold dilution) with incubation for 20 minutes in dark at room temperature. Non-specific Control IgG, clone X63 (cat # M-1249-200) was used as negative control under same conditions (black dashed). Flow cytometry data and results were generated using Orflo MoxiflowTM instrument and protocols. The data demonstrates specific staining of MBP expressed in human neuroblastoma SH-SY5Y cell line using cat # M-1384-50.
Left: Staining of rat hippocampal section with mouse antibody to myelin basic protein (MBP, 1:5,000, green) by Immunohistochemistry. NF-H was co-stained with R-1389-50 (1:2,000, red). The MBP antibody stains oligodendrocyte cell bodies and the myelin sheathes around axons, while the NF-H antibody labels the axons themselves. Right: Western blot analysis of tissue lysates using mouse antibody to MBP (1:20,000, green):  protein standard,  rat brain,  rat spinal cord,  rat sciatic nerve,  mouse brain,  mouse spinal cord. Multiple bands between 14-22 kDa are alternate transcripts of MBP. Other bands are proteolytic fragments of the MBP protein.