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<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D">Hi John,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D">I never got round to either before I left MedImmune, but I was convinced back then that elutriation or A4F would work
better and more robustly for cells and <span class="SpellE">nano</span>-particles than density gradients ever could.<span style="mso-spacerun:yes">
</span><o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D">In elutriation sedimentation is countered by fluid flow in a special centrifuge cell.<span style="mso-spacerun:yes">
</span>I believe that the exact speed is chosen empirically.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D">The new A4F setup with an electric field could also result in a more pure sample since junk with the same hydrodynamic
radius <span style="mso-spacerun:yes"> </span>but a different charge could be separated from the peak of interest by the electric field.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D">Beckman and Wyatt, respectively, can give protocols for these procedures.<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D">Kind Regards,<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D">David<o:p></o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
<p class="MsoNormal"><span style="mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:"Cordia New";color:#1F497D"><o:p> </o:p></span></p>
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<p class="MsoNormal"><b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif";mso-fareast-font-family:"Times New Roman"">From:</span></b><span style="font-size:10.0pt;font-family:"Tahoma","sans-serif";mso-fareast-font-family:"Times New Roman""> RASMB
[mailto:rasmb-bounces@list.rasmb.org] <b>On Behalf Of </b>John Sumida<br>
<b>Sent:</b> Thursday, November 29, 2018 3:36 PM<br>
<b>To:</b> RASMB<br>
<b>Subject:</b> [RASMB] s value and classical sucrose gradient separation<o:p></o:p></span></p>
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<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">Dear RASMB,<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">How would one use the sedimentation value to design a classical sucrose gradient experiment to extract and then purify a species of interest?<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">This question is in relation to the characterization and isolation of nanoparticles.<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">I imagine it would be using the observed sedimentation value obtained experimentally and calculating r to determine the radius at a given buoyance ratio as per<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><img width="307" height="107" id="Picture_x0020_1" src="cid:image001.png@01D487FB.D78892E0"><o:p></o:p></p>
<p class="MsoNormal">Hirst_Cox_Biochem_J_1976_159 (see attached).<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">However, if there is a standard protocol in current use, I would be interested in a reference if appropriate.
<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">Thank you for considering my question.<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal">Best regards,<o:p></o:p></p>
<p class="MsoNormal">John Sumida<o:p></o:p></p>
<p class="MsoNormal">Molecular Analysis Facility<o:p></o:p></p>
<p class="MsoNormal">University of Washington<o:p></o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
<p class="MsoNormal"><o:p> </o:p></p>
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