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<p>And bracket them with as many multiples of blot times and offsets as you have sample and patience to allow!<br>
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<div style="font-family:Tahoma; font-size:13px"><b>Cheers,</b></div>
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<div style="font-family:Tahoma; font-size:13px"><i>Valorie<br>
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<div style="font-family:Tahoma; font-size:13px"><i>Valorie Bowman</i><br>
<br>
vdb@purdue.edu<br>
765-494-5643<br>
EM Facility Laboratory Manager/<br>
Senior Research Electron Microscopist<br>
Purdue Cryo EM Facility<br>
Purdue University</div>
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<div id="divRplyFwdMsg" dir="ltr"><font style="font-size:11pt" color="#000000" face="Calibri, sans-serif"><b>From:</b> 3dem <3dem-bounces@ncmir.ucsd.edu> on behalf of David Michael Belnap <David.Belnap@utah.edu><br>
<b>Sent:</b> Monday, February 26, 2018 11:57 AM<br>
<b>To:</b> Sanchaita Das<br>
<b>Cc:</b> 3dem Scripps<br>
<b>Subject:</b> Re: [3dem] Ice thickness</font>
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<div>Sanchaita,
<div class=""><br class="">
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<div class="">"There is no royal road to [success in cryo-EM].” :-)</div>
<div class=""><br class="">
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<div class="">I would start with settings that have been successful with other samples on that Vitrobot.</div>
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<div class="">David</div>
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<blockquote type="cite" class="">
<div class="">On Feb 26, 2018, at 9:09 AM, Sacha De Carlo <<a href="mailto:sachadecarlo@yahoo.com" class="">sachadecarlo@yahoo.com</a>> wrote:</div>
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<div class="">
<div class="">Unfortunately the molecular weight of the complex alone is useless to determine “proper” settings for the vitrobot. Buffer composition, sample concentration, etc would be required to give you an educated guess ...
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<div class="">Cheers</div>
<div class="">Sacha<br class="">
<br class="">
<br class="">
<a href="https://yho.com/footer0" class="">Sent from Yahoo Mail for iPhone</a><br class="">
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<p class="yahoo-quoted-begin" style="font-size:15px; color:#715FFA; padding-top:15px; margin-top:0">
On Monday, February 26, 2018, 5:05 PM, Sanchaita Das <<a href="mailto:sanchaita06@gmail.com" class="">sanchaita06@gmail.com</a>> wrote:</p>
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<div id="yiv4014431603" class="">
<div class="">Dear all,
<div class="">Does anyone have suggestions about what vitrobot settings may work for getting good single particles for a protein that is aproox 300KDa?</div>
<div class="">I know its trial and error but if something might work for smaller proteins would love suggestions.</div>
<div class="">Best,</div>
<div class="">Sanchaita<br class="">
<br class="">
<div class=""><br class="">
I think 99 times and I find nothing.<br class="">
I stop thinking,<br class="">
swim in silence,<br class="">
and the truth comes to me.<br class="">
<br class="">
- Albert Einstein -<br class="">
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