Total protein levels were analyzed in left hemibrains after homogenisation. Briefly, brain tissues were homogenised in 10:1 v/w tissue homogenisation buffer (0.25 M sucrose, 20 mM Tris‐HCl pH 7.4, 1 mM EDTA, 1 mM EGTA; all reagents from Sigma‐Aldrich) supplemented with protease inhibitors (5 μg/ml leupeptin, 5 μg/ml antipain dihydrochloride, 5 μg/ml pepstatin A, 1 mM phenylmethanesulfonyl fluoride, 1 μM E64; all reagents from Sigma‐Aldrich) immediately before homogenisation. The procedure was performed in ice‐cold glass homogenizers with 20 complete strokes of Teflon pestles (Wheaton, DWK Life Sciences, Millville, NJ, US). Aliquots of homogenate were stored at ‐80°C until use.
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Brain Tissue Homogenization for Protein Analysis
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Corresponding Organization : New York University
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Variable analysis
independent variables
- None explicitly mentioned
- Total protein levels
- Tissue homogenization buffer (0.25 M sucrose, 20 mM Tris‐HCl pH 7.4, 1 mM EDTA, 1 mM EGTA)
- Protease inhibitors (5 μg/ml leupeptin, 5 μg/ml antipain dihydrochloride, 5 μg/ml pepstatin A, 1 mM phenylmethanesulfonyl fluoride, 1 μM E64)
- No positive or negative controls were explicitly mentioned.
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