https://eln.excellgen.com/index.php?action=history&feed=atom&title=Transferin_scFc_formulation 2026-10-18T02:35:50Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=Transferin_scFc_formulation&diff=13131&oldid=prev 2014-09-18T20:48:17Z

<p>Created page with &quot;Preparation of Tf–lipoplexes&lt;br&gt;Cationic liposomes (Lip) consisting of dioleoyl trimethyl-&lt;br&gt;ammonium propane (DOTAP) and dioleoyl phosphatidyl-&lt;br&gt;ethanolamine (DOPE) (Avanti...&quot;</p> <p><b>New page</b></p><div>Preparation of Tf–lipoplexes&lt;br&gt;Cationic liposomes (Lip) consisting of dioleoyl trimethyl-&lt;br&gt;ammonium propane (DOTAP) and dioleoyl phosphatidyl-&lt;br&gt;ethanolamine (DOPE) (Avanti Polar Lipids, Alabaster, AL) at&lt;br&gt;a 1:1 molar ratio were prepared either by sonication or ethanol&lt;br&gt;injection as previously described (L. Xu et al., 1997, 1999a,&lt;br&gt;2001). Tf–Lip–DNA was prepared according to the in vivo for-&lt;br&gt;mulation described previously (L. Xu et al., 1999a). Briefly, for&lt;br&gt;a typical preparation with optimized in vivo formulation, 25 ml&lt;br&gt;of Tf (5 mg/ml, iron-saturated holo-transferrin; Sigma, St.&lt;br&gt;Louis, MO) and 50 ml of Lip (2 mM total lipids) plus 75 ml of&lt;br&gt;water were mixed in a polypropylene tube and incubated for&lt;br&gt;5–15 min at room temperature with frequent rocking. Ten mi-&lt;br&gt;crograms of plasmid DNA in 120 ml of 20 mM HEPES buffer,&lt;br&gt;pH 7.4, was added to the tube, mixed immediately and thor-&lt;br&gt;oughly, and incubated for 20 min at room temperature with fre-&lt;br&gt;quent rocking. Thirty microliters of 50% dextrose solution was&lt;br&gt;then added to the tube. The final DNA:lipid:Tf ratio was&lt;br&gt;1:10:12.5 (mg/nmol/mg). The HEPES buffer can be replaced by&lt;br&gt;pure water.&lt;br&gt;<br /> <br /> <br /> <br /> <br /> <br /> [[Category:Transfection]]</div>

Paul.xu