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<p class="MsoNormal">Yes, we use a mix of lipids.<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal"><b>From:</b> AMARSHI MUKHERJEE <amarshimukherjee@gmail.com> <br>
<b>Sent:</b> Friday, October 15, 2021 11:55 AM<br>
<b>To:</b> Dykstra, Holly <Holly.Dykstra@vai.org><br>
<b>Cc:</b> 3dem@ncmir.ucsd.edu<br>
<b>Subject:</b> [External] Re: [3dem] Membrane protein dissociating from nanodisc<o:p></o:p></p>
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<p class="MsoNormal">Hi,<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal">What is the lipid composition of the nanodiscs? If you are using a single lipid (such as PC only) the chance of incorporation of protein is low sometimes and may cause protein dissociation. if you use a mixture of lipids (such as PC/PS
or PC/PE/PS) it may stabilize the protein in nanodiscs more than a single lipid nanodiscs. <o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal">On Fri, Oct 15, 2021 at 8:13 AM Dykstra, Holly <<a href="mailto:Holly.Dykstra@vai.org">Holly.Dykstra@vai.org</a>> wrote:<o:p></o:p></p>
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<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">Hello all,<o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">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" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">Thanks!<o:p></o:p></p>
<p class="MsoNormal" style="mso-margin-top-alt:auto;mso-margin-bottom-alt:auto">-Holly<o:p></o:p></p>
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<p class="MsoNormal">_______________________________________________<br>
3dem mailing list<br>
<a href="mailto:3dem@ncmir.ucsd.edu" target="_blank">3dem@ncmir.ucsd.edu</a><br>
<a href="https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem" target="_blank">https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem</a><o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal">-- <o:p></o:p></p>
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<p class="MsoNormal">Regards,<o:p></o:p></p>
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<p class="MsoNormal"><o:p> </o:p></p>
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<p class="MsoNormal"><b>Amarshi Mukherjee, M.Sc., Ph.D.</b><o:p></o:p></p>
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<p class="MsoNormal">Research Associate<o:p></o:p></p>
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<p class="MsoNormal">Department of Neuroscience<o:p></o:p></p>
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<p class="MsoNormal">Wisconsin Institute for Medical Research (WIMR)<o:p></o:p></p>
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<p class="MsoNormal">1111 Highland Avenue, Madison, WI 53705<o:p></o:p></p>
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<p class="MsoNormal">Mobile: 443-683-6757<o:p></o:p></p>
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<p class="MsoNormal">Email: <a href="mailto:amukherjee29@wisc.edu" target="_blank">
amukherjee29@wisc.edu</a>, <a href="mailto:amarshimukherjee@gmail.com" target="_blank">
amarshimukherjee@gmail.com</a><o:p></o:p></p>
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<p style="background:#FFEB9C"><strong><span style="font-size:10.0pt;font-family:"Calibri",sans-serif;color:#9C6500;background:#FFEB9C">CAUTION</span></strong><strong><span style="font-size:8.5pt;font-family:"yui-tmp",serif;color:#9C6500;background:#FFEB9C">:</span></strong><span style="font-size:10.0pt;color:black"> This
email was sent from outside of the organization (<a href="mailto:amarshimukherjee@gmail.com">amarshimukherjee@gmail.com</a>). Do not click links or open attachments unless you recognize the sender and know the content is safe. If you have any questions, please
contact <a href="mailto:servicedesk@vai.org">ServiceDesk@vai.org</a>.</span><o:p></o:p></p>
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