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Rockland Immunochemicals
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vdac - by Bioz Stars,
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Rockland Immunochemicals
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SAS institute
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Image Search Results
Journal: Scientific Reports
Article Title: The transcriptome of mouse central nervous system myelin
doi: 10.1038/srep25828
Figure Lengend Snippet: ( A ) Scheme illustrating the biochemical purification of a light-weight membrane fraction enriched for myelin by homogenizing mouse brains in 0.32 M sucrose, sequential sucrose density gradient centrifugation, and osmotic shocks. Myelin accumulates at the interface between 0.32 M and 0.85 M sucrose. ( B ) Immunoblot analysis of myelin-enriched fractions and equal amounts of brain lysate to compare the abundance of marker proteins for compact myelin (PLP/DM20, MBP), non-compact myelin (CNP, SIRT2), the oligodendroglial nuclear/cytoplasmic compartment (OLIG2), astrocytes (GFAP), microglia (AIF1/IBA1), neuronal plasma membrane (GPM6A), axonal microtubules (TUBB3/TUJ1), and mitochondria (VDAC). Blot represents three biological replicates (male c57Bl6/N mice, age P75). Note that myelin markers were enriched in purified myelin while markers of other cellular sources were reduced. ( C ) Heatmap displaying reverse CT values from qRT-PCRs for three markers each specific for myelin, migroglia, neurons and astrocytes performed on myelin biochemically purified from the brains of 4 individual mice (M1–4) compared to the respective brain lysates (BL1–4) at six month of age.
Article Snippet: Antibodies were specific for PLP/DM20 (A431 , 1:5000), MBP (Dako A0623, 1:500), CNP (Sigma C 5922, 1:1000), SIRT2 (Abcam ab67299, 1:500), OLIG2 (DF308 , 1:200, kindly provided by J. Alberta and C. Stiles, Boston, MA, USA) GFAP (Novocastra NCL-GFAP-GA5, 1:500), AIF (also termed IBA1; Abcam ab107159, 1:500), GPM6A (#24924 , 1:1000), TUBB3 (also termed TUJ1; Covance MMS-435P, 1:1000), and
Techniques: Purification, Membrane, Gradient Centrifugation, Western Blot, Marker, Clinical Proteomics