Isolation of low-n amyloid β-protein oligomers from cultured cells, CSF, and brain.
Recent data suggest that soluble, non-fibrillar assemblies of the amyloid β-protein (Aβ) may mediate the synaptic deficits that characterize the early stages of Alzheimer's disease. Consequently, much effort has been expended in isolating and studying a variety of different Aβ assemblies. Here, we describe the use of immunoprecipitation/western blotting and size exclusion chromatography/western blotting to characterize Aβ present in conditioned medium from cultured cells, human cerebrospinal fluid, and human cortex extracted with aqueous buffer, detergent, and formic acid.
Shankar
Ganesh M
GM
Center for Neurologic Diseases, Brigham & Women's Hospital, Harvard Medical School, Boston, MA, USA.
Welzel
Alfred T
AT
McDonald
Jessica M
JM
Selkoe
Dennis J
DJ
Walsh
Dominic M
DM
D016229:Amyloid beta-Peptides / Q000134:cerebrospinal fluid* / Q000302:isolation & purification*; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D002850:Chromatography, Gel; D017077:Culture Media, Conditioned / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D047468:Immunoprecipitation; D066298:In Vitro Techniques
D016229:Amyloid beta-Peptides; D017077:Culture Media, Conditioned