[3dem] Hexamer dissociate while freezing Cryo-EM grids

[Ed Morris]

???Dear Umar

One thing that could be worth checking is to image a range of ice thicknesses in your existing cryo grids. If you are just imaging in the thinnest ice it may be that you have excluded your hexamer and are just seeing smaller complexes. In that case the hexamers may be hiding in thicker ice.

This has happened to me in the past and I initially mistakenly concluded that large particles seen in negative stain were being replaced by subcomplexes when I went I imaged in cryo. On checking the thicker ice I realised that this was not in fact the case.

Best wishes

Ed Morris







________________________________
From: 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of Umar Farook <umarfarook12 at gmail.com>
Sent: 04 June 2020 20:24
To: 3dem at ncmir.ucsd.edu
Subject: [3dem] Hexamer dissociate while freezing Cryo-EM grids

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Dear All,

I would like to ask the community for suggestions on how to overcome the issue of hexamer dissociating while freezing Cryo-EM grids, the negative staining seems to be perfect hexamer at 0.1 mg/ml concentration. But the sample seems to fall apart at 1.5 mg/ml in Cryo freezing, please advise, thank you.

Best Regards,
Umar

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