Bitan:HFIP Film Preparation: Difference between revisions
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<p class=MsoNormal><span style='font-size: | <p class=MsoNormal><span style='font-size:14.0pt;color:#993300'><b>Preparation | ||
1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) film of Aβ sample<o:p></o:p | of 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) film of Aβ sample</b></span><span | ||
style='font-size:14.0pt'><o:p></o:p></span></p> | |||
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<p class=MsoNormal>1) Place HFIP container on ice in the fume hood to pre-chill | <table border=1 cellspacing=0 cellpadding=0 style='border-collapse:collapse; | ||
the HFIP, or just put it in freezer for 10 min. Also it is better to pre-chill | border:none;mso-border-top-alt:solid windowtext .5pt;mso-border-bottom-alt: | ||
solid windowtext .5pt;mso-padding-alt:0in 5.4pt 0in 5.4pt'> | |||
<tr> | |||
<td width=449 valign=top style='width:449.1pt;border-top:solid windowtext .5pt; | |||
border-left:none;border-bottom:solid windowtext .5pt;border-right:none; | |||
padding:0in 5.4pt 0in 5.4pt'> | |||
<p class=MsoNormal>1) Place HFIP container on ice in the fume hood to | |||
pre-chill the HFIP, or just put it in freezer for 10 min. Also it is better | |||
to pre-chill all the tubes.<o:p></o:p></p> | |||
<p class=MsoNormal>2) Add the pre-chilled HFIP to lyophilized peptide | |||
(200ug/40uL). Be quickly, because the HFIP will evaporate very fast.<o:p></o:p></p> | |||
<p class=MsoNormal>3) Incubate the samples at room temperature for 30 min. | |||
Some peptides may require 5 min of sonication for dissolution before | |||
incubation.<o:p></o:p></p> | |||
<p class=MsoNormal>4) If you need divide the sample solution, cool down the | |||
tubes again and divide 10 uL per PCR tube or regular tube. Leave the caps of | |||
tubes open for several hours or overnight to evaporate the most of HFIP. It’s | |||
better to cover the tubes with a piece of kimwipe tissue paper to prevent the | |||
dust.<o:p></o:p></p> | |||
<p class=MsoNormal>5) Speed vacuuming the rest small amount of HFIP.<o:p></o:p></p> | |||
<p class=MsoNormal>6) Para film closed at -20 degree to store. Good for a few | |||
months.<o:p></o:p></p> | |||
</td> | |||
</tr> | |||
</table> | |||
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Revision as of 23:51, 22 November 2009
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<p class=MsoNormal><span style='font-size:14.0pt;color:#993300'><b>Preparation of 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) film of Aβ sample</b></span><span style='font-size:14.0pt'><o:p></o:p></span></p>
<p class=MsoNormal style='tab-stops:159.2pt'><span style='mso-tab-count:1'> </span></p>
<table border=1 cellspacing=0 cellpadding=0 style='border-collapse:collapse;
border:none;mso-border-top-alt:solid windowtext .5pt;mso-border-bottom-alt: solid windowtext .5pt;mso-padding-alt:0in 5.4pt 0in 5.4pt'> <tr> <td width=449 valign=top style='width:449.1pt;border-top:solid windowtext .5pt; border-left:none;border-bottom:solid windowtext .5pt;border-right:none; padding:0in 5.4pt 0in 5.4pt'> <p class=MsoNormal>1) Place HFIP container on ice in the fume hood to pre-chill the HFIP, or just put it in freezer for 10 min. Also it is better to pre-chill all the tubes.<o:p></o:p></p> <p class=MsoNormal>2) Add the pre-chilled HFIP to lyophilized peptide (200ug/40uL). Be quickly, because the HFIP will evaporate very fast.<o:p></o:p></p> <p class=MsoNormal>3) Incubate the samples at room temperature for 30 min. Some peptides may require 5 min of sonication for dissolution before incubation.<o:p></o:p></p> <p class=MsoNormal>4) If you need divide the sample solution, cool down the tubes again and divide 10 uL per PCR tube or regular tube. Leave the caps of tubes open for several hours or overnight to evaporate the most of HFIP. It’s better to cover the tubes with a piece of kimwipe tissue paper to prevent the dust.<o:p></o:p></p> <p class=MsoNormal>5) Speed vacuuming the rest small amount of HFIP.<o:p></o:p></p> <p class=MsoNormal>6) Para film closed at -20 degree to store. Good for a few months.<o:p></o:p></p> </td> </tr>
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