Team:TU Darmstadt/protocols/SDS-Page

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<font size="3" color="#F0F8FF" face="Arial regular">
<font size="3" color="#F0F8FF" face="Arial regular">
<p text-aligne:left style="margin-left:50px; margin-right:50px">
<p text-aligne:left style="margin-left:50px; margin-right:50px">
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<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
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<br>
 +
<B>Equipment<br></B>
 +
<div align="left" style="margin-left:60px; margin-right:50px">
 +
<ul>
 +
<li class=list1>- Two glass plates</li>
 +
<li class=list1>- Gel caster</li>
 +
<li class=list1>- Comb</li>
 +
<li class=list1>- VWR Power Source 300V</li>
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</ul>
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</div>
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</font>
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</p>
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<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
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<br>
 +
<B>Chemicals & consumables<br></B>
 +
<div align="left" style="margin-left:60px; margin-right:50px">
 +
<ul>
 +
<li class=list1>- SDS</li>
 +
<li class=list1>- Rotiphorese® (30%)</li>
 +
<li class=list1>- Tris HCl</li>
 +
<li class=list1>- Glycine</li>
 +
<li class=list1>- TEMED</li>
 +
<li class=list1>- APS</li>
 +
<li class=list1>- Aqua dest.</li>
 +
<li class=list1>- Isopropyle alcohol </li>
 +
 +
</ul>
 +
</div>
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</font>
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</p>
 +
 +
<p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
 +
<br>
 +
<B>Buffers<br></B>
 +
<div align="left" style="margin-left:60px; margin-right:50px">
 +
<ul>
 +
<li class=list1>- Separation buffer (18.17 g Tris, 0.40 g SDS, 90 ml Aqua dest., pH = 8.8)</li>
 +
<li class=list1>- Rotiphorese® (30%)</li>
 +
<li class=list1>- Tris HCl</li>
 +
<li class=list1>- Glycine</li>
 +
<li class=list1>- TEMED</li>
 +
<li class=list1>- APS</li>
 +
<li class=list1>- Aqua dest.</li>
 +
<li class=list1>- Isopropyle alcohol </li>
 +
 +
</ul>
 +
</div>
 +
</font>
 +
</p>
 +
<br>
 +
<p text-aligne:left style="margin-left:50px; margin-right:50px"><font size="4.5" color="#F0F8FF" face="Arial regular">
 +
<B> Procedure<br></B></font></p>
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<p text-aligne:left style="margin-left:60px; margin-right:60px"><font size="3" color="#F0F8FF" face="Arial regular">
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 +
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<div align="left" style="margin-left:30px; margin-right:50px">
 +
<ol>
 +
<li>prepare the separating gel and fill it into the chamber </li>
 +
<li>pour 1 mL isopropyle alcohol on the top of the gel to destroy air bubbles and prevent dehydration </li>
 +
<li>discard the isoprpyl alcohol and pour the prepared stacking gel</li>
 +
<li>stick in the comb</li>
 +
<li>store the gel in wet cloth (to prevent dehydration) at 4°C</li>
 +
</ol>
 +
</div>
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</font></p>

Revision as of 18:39, 4 October 2013







Labbook | Matierals | Protocols

Protocols


Equipment

  • - Two glass plates
  • - Gel caster
  • - Comb
  • - VWR Power Source 300V


Chemicals & consumables

  • - SDS
  • - Rotiphorese® (30%)
  • - Tris HCl
  • - Glycine
  • - TEMED
  • - APS
  • - Aqua dest.
  • - Isopropyle alcohol


Buffers

  • - Separation buffer (18.17 g Tris, 0.40 g SDS, 90 ml Aqua dest., pH = 8.8)
  • - Rotiphorese® (30%)
  • - Tris HCl
  • - Glycine
  • - TEMED
  • - APS
  • - Aqua dest.
  • - Isopropyle alcohol


Procedure

  1. prepare the separating gel and fill it into the chamber
  2. pour 1 mL isopropyle alcohol on the top of the gel to destroy air bubbles and prevent dehydration
  3. discard the isoprpyl alcohol and pour the prepared stacking gel
  4. stick in the comb
  5. store the gel in wet cloth (to prevent dehydration) at 4°C