<html><head><meta http-equiv="content-type" content="text/html; charset=utf-8"></head><body dir="auto">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. <div><br></div><div>Cheers</div><div>Jacopo <br><br><div id="AppleMailSignature" dir="ltr"><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></div><div dir="ltr"><br>On 15 Oct 2021, at 15:13, Dykstra, Holly <<a href="mailto:Holly.Dykstra@vai.org">Holly.Dykstra@vai.org</a>> wrote:<br><br></div><blockquote type="cite"><div dir="ltr">
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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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