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<p>what form of nano disc are you using?</p>
<p>We do a fair amount of work on the SMALP system (we developed it) so might be able to help</p>
<p>Tim</p>
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Tim R. Dafforn PhD FIET<br>
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<div class="PlainText">Professor of Biotechnology<br>
University of Birmingham<br>
Edgbaston<br>
Birmingham<br>
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+44 (0) 121 414 5881</div>
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<div id="divRplyFwdMsg" dir="ltr"><font face="Calibri, sans-serif" style="font-size:11pt" color="#000000"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of jacopo.marino@psi.ch <jacopo.marino@psi.ch><br>
<b>Sent:</b> 15 October 2021 14:39:07<br>
<b>To:</b> Dykstra, Holly<br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> Re: [3dem] Membrane protein dissociating from nanodisc</font>
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<div>Hi - one would expect that membrane proteins embedded into nanodiscs would suffer less than detergent-embedded proteins, but that’s not a rule. The question is simple: is it faster to live with your small amount of particles and compensate with collecting
many more datasets, or screening for freezing conditions where you have less dissociation ? You might find that areas with thicker ice on your grid have more intact particles, but likely you don’t want to collect from there.
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<div>Cheers</div>
<div>Jacopo <br>
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<pre class="moz-signature" cols="72"><font face=".SFUIDisplay"><span style="white-space: normal; background-color: rgba(255, 255, 255, 0);">Dr. Jacopo Marino
Laboratory of Biomolecular Research
Paul Scherrer Institute
OFLB/005
5232 </span></font></pre>
<pre class="moz-signature" cols="72"><font face=".SFUIDisplay"><span style="white-space: normal; background-color: rgba(255, 255, 255, 0);">Villigen PSI, Switzerland</span></font></pre>
<pre class="moz-signature" cols="72"><font face=".SFUIDisplay"><span style="white-space: normal; background-color: rgba(255, 255, 255, 0);">tel: <a href="tel:+41%2056%20310%205777" dir="ltr" x-apple-data-detectors="true" x-apple-data-detectors-type="telephone" x-apple-data-detectors-result="50/0" style="-webkit-text-decoration-color: rgba(0, 0, 0, 0.258824);">+41 56 310 5777</a></span></font></pre>
<pre class="moz-signature" cols="72"><font face=".SFUIDisplay"><span style="white-space: normal; background-color: rgba(255, 255, 255, 0);">e-mail: <a class="moz-txt-link-abbreviated" href="mailto:jacopo.marino@psi.ch">jacopo.marino@psi.ch</a></span></font></pre>
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On 15 Oct 2021, at 15:13, Dykstra, Holly <<a href="mailto:Holly.Dykstra@vai.org">Holly.Dykstra@vai.org</a>> wrote:<br>
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<p class="MsoNormal">Hello all,<o:p></o:p></p>
<p class="MsoNormal">I am working on a single-TM membrane protein in nanodisc. Upon freezing grids, I am observing protein dissociation from the nanodisc. A small percentage of the total particle population is intact protein-nanodisc complex but most of the
particles have dissociated. I believe this is happening at the point of freezing as negative stain shows the intact protein-nanodisc complex. Has anyone else experienced this problem or does anyone have any suggestions or advice?<o:p></o:p></p>
<p class="MsoNormal">Thanks!<o:p></o:p></p>
<p class="MsoNormal">-Holly<o:p></o:p></p>
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