I need to purify a class of synaptosomes (so I can start from a synaptosome preparation with conventional methods) by affinity tag. The tag would be 3xFLAG -TEV-HisTag and is in the extracellular part of a TM protein localized on the postsynaptic site.
Has anybody any experience in puryfing synaptosomes with antobodies or related?
Thank you in advance.
Dear Rafael
No, it's not a transgenic, I will be infecting the hippocampus with a AAV vector
Then yes I would go ahead but I would not discard any membranous fraction to check if your protein goes with your "synaptosomal" preparation or with the other membranes or both (and in what amount each). Do you plan to use detergent to make your protein soluble? This may be a crucial step.
No detergent, we are targeting the extracellular epitope for pull down, then we are just sending the sample to mass spectrum analysis
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