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Dear Nikhil,
First - I like your email address...
Out of the list you ask about, I have used just PSD95 antibodies. They worked very well in samples fixed with methanol, but did not work whatsoever in PFA fixed samples. So, at least in my case, this was a necessity. At the light-microscope level I could not see any obvious problem with the methanol fixed sample, when comparing other markers I had used (like presynaptic proteins) in methanol and PFA fixed samples. Notice that when hydrating the cultures after the methanol fixation step, the solutions bubble quite a lot. Still, the end results were as expected, and quite nice to behold Hope this helps.
Daniel

Daniel Gitler, Senior Lecturer
Department of Physiology and Cell Biology
Faculty of Health Sciences
Ben-Gurion University of the Negev
Beer-Sheva 84105
Israel

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From: Confocal Microscopy List [[log in to unmask]] on behalf of nikhil pandya [[log in to unmask]]
Sent: Wednesday, October 22, 2014 10:42 PM
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Subject: Methanol fixation for primary cultures neurons.

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Greetings!
A bit of Neuroscience specific question.

I have been testing antibodies for Post synaptic density proteins like
PSD95 and AMPA and NMDA receptor in primary hippocampal neurons. I am
finding it difficult to get good labelling in the PSD. Is it recommended to
use methanol to fix and stain PSD proteins in general? Are there any
disadvantages to using methanol instead of PFA fixation?

--
Nikhil Janak Pandya.
E mail: [log in to unmask]
Contact no: +316631317506