Tissue Homogenization and Fractionation for Protein Analysis

Tissues were prepared as previously described^{29 (link)}. Briefly, the tissues were dissected into small pieces and homogenized at 4 °C using the Precellys 24 homogenizer in PBS with protease and phosphatase inhibitors (Roche,Indianapolis, Indiana). For fractionation, brain tissue was homogenized in a teflon dounce grinder on ice in PBS with protease and phosphatase inhibitors (Roche, Indianapolis, Indiana). After homogenization, protein concentration was determined with a Pierce BCA protein assay (Life Technologies, Grand Island, NY).

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McClatchy D.B., Martínez-Bartolomé S., Gao Y., Lavallée-Adam M, & Yates JR I.I.I. (2020). Quantitative analysis of global protein stability rates in tissues. Scientific Reports, 10, 15983.

Publication 2020

Precellys 24 protease and phosphatase inhibitors Pierce BCA protein assay

Assay Brain Fractionation Inhibitors Phosphatase Protease Protein Protein a Teflon Tissue

Corresponding Organization :

Other organizations : Scripps Research Institute, University of Illinois at Chicago

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Variable analysis

independent variables

Tissue preparation method (dissection into small pieces and homogenization at 4 °C using the Precellys 24 homogenizer)

dependent variables

Protein concentration

control variables

PBS with protease and phosphatase inhibitors (Roche, Indianapolis, Indiana) used for homogenization
Temperature (4 °C) for homogenization

controls

No positive or negative controls were explicitly mentioned in the provided information.

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