User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/04/27: Difference between revisions

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{|{{table}} width="800"
{|{{table}} width="800"
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">RII Overlay</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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*RIPA BUFFER
*RIPA BUFFER (100 mL)
 
**50 mM Tris-HCl (5 mL 1 M Tris pH 7.4)
**Adjust to pH 7.4
**150 mM NaCl (0.87 g NaCl)
** 1 mM EDTA (37.22 mg EDTA)
**1% Triton x-100 (1 mL Tritox x-100)
**1% Sodium deoxycholate
**0.1% SDS (1 g SDS)
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==Materials & Methods==
==Materials & Methods==
===Materials===
===Materials===
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   <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">
   <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">
</a>
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</html> [[Media:FILE.xls| Excel TITLE]]
</html> [[Media:FILE.xls| Excel TITLE]]-->
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__NOTOC__
__NOTOC__

Revision as of 01:32, 11 June 2010

<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>

RII Overlay <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Summary

  • RII overlay
    • Activate membrane in EtOH 100% for 15 sec.
    • Rinse with H2O
    • Block in Blocking buffer for 3 hours
      • Blocking buffer (5% Skim milk, 0.1% BSA, 0.0001% Sodium Azide in PBS)


  • RIPA BUFFER (100 mL)
    • 50 mM Tris-HCl (5 mL 1 M Tris pH 7.4)
    • Adjust to pH 7.4
    • 150 mM NaCl (0.87 g NaCl)
    • 1 mM EDTA (37.22 mg EDTA)
    • 1% Triton x-100 (1 mL Tritox x-100)
    • 1% Sodium deoxycholate
    • 0.1% SDS (1 g SDS)


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