Does anyone have experience in purifying plasma membrane from dorsal root ganglion? And even better, neuronal plasma membrane? We want to quantitate receptor membrane insertion within the ganglionic neurons.
Hi Sean,
Well, I'm not a specialist in DRG purification, I only work on full mouse brain to isolate PM and Microsomal fractions,but you can use a subcellular fractionation protocol (multiple centrifugation). It works very well for me.
However, there is this article that describe a good method for DRG neurons.
https://proteomesci.biomedcentral.com/articles/10.1186/1477-5956-7-41
Thanks for the link, Sebastien! I had heard about two-phase purification many years ago, but had completely forgotten about it. I think that it will be worth a try.
Best wishes,
Sean
Well, I'm not an expert at membrane purification . But here is my idea of bypassing it. If you know the speed of endocytosis of your receptor under a given condition, and can block the insertion of receptors on PM under the same condition with a specific temporal control, the calculation will be much easier.
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