For the analyses of the relative amounts of Bid and tBid in P4 vs. P7 cerebellum following exposure to experimental conditions, tissue was fractionated, to separate cytosolic and mitochondrial contents. Full-length Bid is a cytosolic protein, and was assessed in cytosolic fractions, while tBid moves to the mitochondrial membrane and was accessed in mitochondrial fractions. For this procedure, cerebellae were harvested immediately after termination of experimental treatment. Tissues were homogenized and separated into fractions using the Mitochondrial/Cytosol Fractionation Kit and protocol provided by BioVision Inc. (Mountain View, CA; catalog #K256), as described in detail previously (e.g., Heaton et al., 2011 (link); 2013 (link)).
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Cerebellum Fractionation for Bid and tBid
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Cerebellum
Cytosol
Experimental treatment
Fractionation
Mitochondrial
Mitochondrial membrane
Protein
Tissues
Corresponding Organization :
Other organizations : Florida College, University of Florida, Addiction Research Foundation, Allen Institute for Brain Science
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Variable analysis
independent variables
- Experimental conditions
- Relative amounts of Bid and tBid in P4 vs. P7 cerebellum
- Tissue fractionation to separate cytosolic and mitochondrial contents
- Cerebellae harvested immediately after termination of experimental treatment
- Tissues homogenized and separated into fractions using the Mitochondrial/Cytosol Fractionation Kit and protocol provided by BioVision Inc.
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