Mouse brain dissection and homogenization

Xu-Qiao Chen; Utpal Das; Gooho Park; William C. Mobley

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Mouse brain dissection and homogenization

XC Xu-Qiao Chen

UD Utpal Das

GP Gooho Park

WM William C. Mobley

This method is extracted from research article: Alzheimers Res Ther, Mar 2021

Normal levels of KIF5 but reduced KLC1 levels in both Alzheimer disease and Alzheimer disease in Down syndrome: evidence suggesting defects in anterograde transport

DOI: 10.1186/s13195-021-00796-6

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Brains were dissected using established landmarks. For Dp16 and age-matched 2N mice, the forebrain was isolated by first removing the olfactory bulbs; cerebellum and brain stem were then removed via a cut anterior to the cerebellum and extending through the posterior midbrain. Dissected forebrain was then immediately frozen in dry ice. The whole brains of J20 and APP knockout mice and age-matched wild-type mice, minus olfactory bulbs, were also dissected and frozen in dry ice. The brain, kidney, and liver were dissected from 2N adult mice to evaluate antibody specificity. Brain tissues were homogenized via a Dounce homogenizer in RIPA buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, and protease inhibitor cocktail) (1 ml for each hemibrain) and rotated at 4 °C for 30 min. All lysates were centrifuged (12,000 rpm for 15 min at 4 °C) to produce supernatants, and the protein content of supernatants was measured by Bradford assay.

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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