Biochemical fractionation of brain tissue for studies of receptor distribution and trafficking.
An important tool for studying the regulation of synapses is a rapid and reliable means of separating synaptic and intracellular proteins. This unit presents a technique for analysis of brain tissue which relies on differential centrifugation to separate proteins present at synaptic sites from those found in intracellular cytoplasmic and vesicular pools. The method is efficient in that only small amounts of tissue, such as might be obtained from a small region of a rodent brain, are required. It is reproducible and, in conjunction with immunoblot or immunoprecipitation techniques, can produce reliable quantitative data. The protocol will be of interest to those conducting a variety of different studies related to the localization and trafficking of brain receptors and signaling molecules.
Hallett
Penelope J
PJ
MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts, USA.
Collins
Tiffany L
TL
Standaert
David G
DG
Dunah
Anthone W
AW
D000818:Animals; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry / Q000502:physiology; D002458:Cell Fractionation / Q000379:methods*; D021381:Protein Transport / Q000502:physiology*; D051381:Rats; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D013569:Synapses / Q000378:metabolism
D011956:Receptors, Cell Surface