Synaptosomal and Cytosolic Fractionation from Brain

To isolate synaptosomal and cytosolic fractions from brain regions of interest, the Syn-PER Synaptic Protein Extraction Reagent (Thermo Fisher Scientific, Waltham, MA, USA) was used. The isolated synaptosomes were lysed in the N-PER Neuronal Protein Extraction Reagent (Thermo Fisher Scientific) prior to any downstream applications, and all extraction reagents were supplemented with Pierce Protease and Phosphatase Inhibitor Mini Tablets (Thermo Fisher Scientific), as previously described [14 (link)]. We previously validated the efficacy of the synaptosomal/cytosolic extraction method by probing for proteins that are enriched in the corresponding fractions [14 (link)].

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Qvist J.S., Scherma M., Jayaram-Lindström N., Fratta W., Kandel D.B., Kandel E.R., Fadda P, & Melas P.A. (2022). Synaptoproteomic Analysis of the Prefrontal Cortex Reveals Spatio-Temporal Changes in SYNGAP1 Following Cannabinoid Exposure in Rat Adolescence. International Journal of Molecular Sciences, 24(1), 698.

Publication 2022

Syn-PER Synaptic Protein Extraction Reagent N-PER Neuronal Protein Extraction Reagent Pierce Protease and Phosphatase Inhibitor Mini Tablets

Brain Cytosolic Neuronal Phosphatase Protease Proteins Synaptosomal

Corresponding Organization : Karolinska University Hospital

Other organizations : Columbia University, University of Cagliari

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Variable analysis

independent variables

Synaptosomal and cytosolic fractions from brain regions of interest

dependent variables

Protein composition and enrichment of synaptosomal and cytosolic fractions

control variables

Extraction reagents supplemented with Pierce Protease and Phosphatase Inhibitor Mini Tablets

controls

Positive control: Proteins known to be enriched in the corresponding synaptosomal and cytosolic fractions
Negative control: Not explicitly mentioned

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