User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/06/28: Difference between revisions

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===Protein work===
===Protein work===
* HeLa cell lysis as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/08|08April2010]]
* HeLa cell lysis as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/08|08April2010]]
** 500 μL RIPA buffer for T25 flask (2 more T75 flasks were ordered)
* Protein determination as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|21June2010]]
* Protein determination as described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|21June2010]]
** HeLa
** HeLa
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** hTERT ASM Samples
** hTERT ASM Samples
*cAMP precipitation
*cAMP precipitation
* SDS-PAGE
**Everything was put on beads corrected to the lowest concentration, to at least get similar relative amounts (see appendix)
** RII Overlay
** Beads were incubate ON @ 4 °C
** Silver / Coomassie staining
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** Immunoblot
*** AKAP250
*** AKAP95
*** AKAP450
===RNA work===
* Preparing cDNA
* Real time qPCR


==Materials & Methods==
==Materials & Methods==
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===Same actions===
===Same actions===
===Related topics===
===Related topics===
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==Attachment==
==Attachment==
<html>
<html>
<a href="http://openwetware.org/wiki/Image:HTERT_Donor_12_ht31_160310.xls">
<a href="http://openwetware.org/wiki/Image:Bradford_28062010.xls">
   <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>
</a>
</html> [[Media:FILE.xls| Excel TITLE]]
</html> [[Media:Bradford_28062010.xls| Bradford results]]
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Revision as of 09:22, 28 June 2010

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Summary

Protein work

  • HeLa cell lysis as described 08April2010
    • 500 μL RIPA buffer for T25 flask (2 more T75 flasks were ordered)
  • Protein determination as described 21June2010
    • HeLa
    • HEK293
    • hTERT ASM Samples
  • cAMP precipitation
    • Everything was put on beads corrected to the lowest concentration, to at least get similar relative amounts (see appendix)
    • Beads were incubate ON @ 4 °C

Attachment

<html> <a href="http://openwetware.org/wiki/Image:Bradford_28062010.xls"> 

  <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">

</a> </html> Bradford results


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