<div dir="ltr"><div dir="ltr"><div dir="auto">Hi Sasha,<div dir="auto"><br></div><div dir="auto">I found Holger Stark's GraFix method very helpful for Cryo-EM work with transcription factor and polymerase complexes, which may be similar to your situation. Holger's paper covers the technique, and you can also look at my and other publications that benefited from this technique; some links below. Happy to discuss further.</div><div dir="auto"><br></div><div>Good luck!</div><div>Brian</div><div dir="auto"><br></div><div dir="auto">Holger's paper: <a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__pubmed.ncbi.nlm.nih.gov_20887855_&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=DNcmcw2ienZ_yQ624pixtXSpjjgFaZqmD6xLZnsRU9YZrY_KqbdpPTo0VRc3A0tf&s=2kWdQFZQkdFBxBbrPawQNBJRSI9MJihWzJeGVTWMQyM&e=">https://pubmed.ncbi.nlm.nih.gov/20887855/</a> <li style="color:rgb(33,33,33);font-family:BlinkMacSystemFont,-apple-system,"Segoe UI",Roboto,Oxygen,Ubuntu,Cantarell,"Fira Sans","Droid Sans","Helvetica Neue",sans-serif;font-size:16px;box-sizing:inherit;line-height:1.5;margin:0px 1rem 0px 0px;display:inline-block;padding:0px"><span class="gmail-identifier gmail-pubmed" style="box-sizing:inherit"><span class="gmail-id-label" style="box-sizing:inherit">PMID: </span><span class="gmail-current-id" title="PubMed ID" style="box-sizing:inherit">20887855</span></span></li><span style="color:rgb(33,33,33);font-family:BlinkMacSystemFont,-apple-system,"Segoe UI",Roboto,Oxygen,Ubuntu,Cantarell,"Fira Sans","Droid Sans","Helvetica Neue",sans-serif;font-size:16px"> </span><li style="color:rgb(33,33,33);font-family:BlinkMacSystemFont,-apple-system,"Segoe UI",Roboto,Oxygen,Ubuntu,Cantarell,"Fira Sans","Droid Sans","Helvetica Neue",sans-serif;font-size:16px;box-sizing:inherit;line-height:1.5;margin:0px 1rem 0px 0px;display:inline-block;padding:0px"><span class="gmail-identifier gmail-doi" style="box-sizing:inherit"><span class="gmail-id-label" style="box-sizing:inherit">DOI: </span><a class="gmail-id-link" target="_blank" rel="noopener" href="https://urldefense.proofpoint.com/v2/url?u=https-3A__doi.org_10.1016_s0076-2D6879-2810-2981005-2D5&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=DNcmcw2ienZ_yQ624pixtXSpjjgFaZqmD6xLZnsRU9YZrY_KqbdpPTo0VRc3A0tf&s=B8fLicxnEuHEAYzu9stkDunWm2FF-WFq8Qeyb7j9AWo&e=" style="box-sizing:inherit;background-color:transparent;color:rgb(32,84,147);text-decoration-line:none;outline:0px;font-size:inherit">10.1016/S0076-6879(10)81005-5</a></span></li><li style="color:rgb(33,33,33);font-family:BlinkMacSystemFont,-apple-system,"Segoe UI",Roboto,Oxygen,Ubuntu,Cantarell,"Fira Sans","Droid Sans","Helvetica Neue",sans-serif;font-size:16px;box-sizing:inherit;line-height:1.5;margin:0px 1rem 0px 0px;display:inline-block;padding:0px"><div dir="auto"></div></li><li style="color:rgb(33,33,33);font-family:BlinkMacSystemFont,-apple-system,"Segoe UI",Roboto,Oxygen,Ubuntu,Cantarell,"Fira Sans","Droid Sans","Helvetica Neue",sans-serif;font-size:16px;box-sizing:inherit;line-height:1.5;margin:0px 1rem 0px 0px;display:inline-block;padding:0px"><br></li><li style="color:rgb(33,33,33);font-family:BlinkMacSystemFont,-apple-system,"Segoe UI",Roboto,Oxygen,Ubuntu,Cantarell,"Fira Sans","Droid Sans","Helvetica Neue",sans-serif;font-size:16px;box-sizing:inherit;line-height:1.5;margin:0px 1rem 0px 0px;display:inline-block;padding:0px"><div dir="auto"></div></li><li style="color:rgb(33,33,33);font-family:BlinkMacSystemFont,-apple-system,"Segoe UI",Roboto,Oxygen,Ubuntu,Cantarell,"Fira Sans","Droid Sans","Helvetica Neue",sans-serif;font-size:16px;box-sizing:inherit;line-height:1.5;margin:0px 1rem 0px 0px;display:inline-block;padding:0px"><br></li></div></div></div>Other work where this technique was applied to stabilize protein interactions for structural work:<div><br><div><a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__www.pnas.org_content_109_6_1949&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=DNcmcw2ienZ_yQ624pixtXSpjjgFaZqmD6xLZnsRU9YZrY_KqbdpPTo0VRc3A0tf&s=jKbp75Nznd_-_8b7WB33uTbKA08IByCB82Zl8JnDdm8&e=">https://www.pnas.org/content/109/6/1949</a> <a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__doi.org_10.1073_pnas.1105266109&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=DNcmcw2ienZ_yQ624pixtXSpjjgFaZqmD6xLZnsRU9YZrY_KqbdpPTo0VRc3A0tf&s=m_2VTcbo96Jfm0eW2VOlsGXlMmZ4xhgpTHatKhe2y8w&e=" style="box-sizing:border-box;text-decoration-line:none;outline:0px;color:rgb(0,90,150);font-family:"Open Sans",HelveticaNeue,"Helvetica Neue",Helvetica,Arial,sans-serif;font-size:12px">https://doi.org/10.1073/pnas.1105266109</a></div><div><a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__www.ncbi.nlm.nih.gov_pmc_articles_PMC4039082_&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=DNcmcw2ienZ_yQ624pixtXSpjjgFaZqmD6xLZnsRU9YZrY_KqbdpPTo0VRc3A0tf&s=OxqvkSEElNWpz5-i49XfosNh2AGTKJ-hxhc4p4NXFik&e=">https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4039082/</a> <a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__www.ncbi.nlm.nih.gov_entrez_eutils_elink.fcgi-3Fdbfrom-3Dpubmed-26retmode-3Dref-26cmd-3Dprlinks-26id-3D24072820&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=DNcmcw2ienZ_yQ624pixtXSpjjgFaZqmD6xLZnsRU9YZrY_KqbdpPTo0VRc3A0tf&s=6h7zV0kLAXdvy-X6UtpJMFalAcmNSRTvPyPO2HR6lTc&e=" target="_blank" rel="noopener noreferrer" style="outline:0px;color:rgb(152,87,53);font-family:arial,helvetica,clean,sans-serif;font-size:13.4792px">Science. 2013 Nov 8; 342(6159): 1238724</a> <span style="color:rgb(0,0,0);font-family:arial,helvetica,clean,sans-serif;font-size:13.4792px;white-space:nowrap">doi: </span><a href="https://urldefense.proofpoint.com/v2/url?u=https-3A__dx.doi.org_10.1126-252Fscience.1238724&d=DwMFaQ&c=-35OiAkTchMrZOngvJPOeA&r=L7-zyQ-04fFCMRqzLIOnx7H0exGZHwIQe_wMPuY600I&m=DNcmcw2ienZ_yQ624pixtXSpjjgFaZqmD6xLZnsRU9YZrY_KqbdpPTo0VRc3A0tf&s=ERkqyGKhvUQa6Hi41SW_P6zIRERnlx62rKVZAY29YY8&e=" target="_blank" rel="noopener noreferrer" style="outline:0px;color:rgb(152,87,53);font-family:arial,helvetica,clean,sans-serif;font-size:13.4792px;white-space:nowrap">10.1126/science.1238724</a><br></div><div><br></div><div><br><div><br></div><div><br></div><div><br></div><div><br></div><div><br></div><div><br><div class="gmail_quote"><div dir="ltr" class="gmail_attr">On Mon, Nov 22, 2021, 5:28 PM SSS Sasha <<a href="mailto:sss_jiandan@outlook.com" target="_blank">sss_jiandan@outlook.com</a>> wrote:<br></div><blockquote class="gmail_quote" style="margin:0px 0px 0px 0.8ex;border-left:1px solid rgb(204,204,204);padding-left:1ex">
<div dir="ltr">
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;font-size:12pt;color:rgb(0,0,0)">
Hi all,</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;font-size:12pt;color:rgb(0,0,0)">
<div> </div>
<div>We are working with two protein complexes that weakly interact with each other. We cannot isolate a stable complex using gel filtration or other similar approaches. We are wondering if it is possible to capture the complex and interaction using crosslinking
reagents. We have seen the use of BS3 in some cryo-EM structures and wondering if you have suggestions and advice on other crosslinking reagents that have been successfully used for high resolution cryo-EM structures, and how to select the best reagent and
condition. </div>
<div> </div>
Thank you very much for your help,<br>
</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;font-size:12pt;color:rgb(0,0,0)">
<br>
</div>
<div style="font-family:Calibri,Arial,Helvetica,sans-serif;font-size:12pt;color:rgb(0,0,0)">
Shasha</div>
</div>
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