User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization

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* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18| ELISA]] ([[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25|Coating buffer]])
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18| ELISA]] ([[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25|Coating buffer]])
* Co-immunoprecipitation [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/23| Part I]],  [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/24| Part II]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25| Part III]]
* Co-immunoprecipitation [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/23| Part I]],  [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/24| Part II]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25| Part III]]
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* cAMP-precipitarion [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/08|Part I]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/09#Washing_beads:_cAMP_precipitation|Part II]]
+
* cAMP-precipitation [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/08|Part I]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/09#Washing_beads:_cAMP_precipitation|Part II]]
** [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/27#Summary|RIPA Buffer (Berlin)]]
** [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/27#Summary|RIPA Buffer (Berlin)]]
-
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/24|Pierce determination]] (protein conc.)
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** [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/08#Discussion|RIPA Buffer (Groningen)]]
 +
* Protein concentration determination: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/24|Pierce determination]] & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|Bradford assay]]
 +
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/25#Coomassie_staining|Coomassie staining]]
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/01| Western blot]]
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/01| Western blot]]
** [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/27#Stripping_blot|Stripping blot]]
** [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/27#Stripping_blot|Stripping blot]]
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* Presentation [[Image:PRESENTATION_POWERPOINT_August2010.ppt|August]]
====Convenient Links====
====Convenient Links====
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</a>
</a>
</html> [[Media:BLANCO_PROTEIN_SIZE.xls| Blanc for protein size on gel/membrane (non-Linear regression)]]
</html> [[Media:BLANCO_PROTEIN_SIZE.xls| Blanc for protein size on gel/membrane (non-Linear regression)]]
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<html>
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<a href="http://openwetware.org/wiki/Image:BLANCO_IF_COPY_ME_BEFORE_USE.xls.xls">
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  <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50">
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</a>
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</html> [[Media:BLANCO_IF_COPY_ME_BEFORE_USE.xls.xls| Blanc form IF coverslips]]
==Table of contents==
==Table of contents==
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|style="background:orange" |
|style="background:orange" |
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<b>February</b>
* 10/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/10| Splitting ASM cells]]
* 10/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/10| Splitting ASM cells]]
* 12/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/12| Counting cells]]
* 12/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/12| Counting cells]]
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* 25/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25| Co-immunoprecipitation part III & Viability staining]]  
* 25/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25| Co-immunoprecipitation part III & Viability staining]]  
* 26/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/26| ELISA, counting/splitting cells]]
* 26/02/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/26| ELISA, counting/splitting cells]]
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<b>March</b>
* 01/03/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/01| Western blot]]
* 01/03/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/01| Western blot]]
* 03/03/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/03| Co-IP day I, GAPDH, VASP, Stop Stimulation HT4]]  
* 03/03/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/03| Co-IP day I, GAPDH, VASP, Stop Stimulation HT4]]  
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* 31/03/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/31|Building smoke machine, cell splitting]]
* 31/03/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/31|Building smoke machine, cell splitting]]
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<b>April</b>
* 01/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/01|Creating RII overlay protocol]]
* 01/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/01|Creating RII overlay protocol]]
* 06/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/06|Putting cells to S<sup>0</sup> and splitting cells]]
* 06/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/06|Putting cells to S<sup>0</sup> and splitting cells]]
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*29/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/29|SDS-PAGE& Finalizing RII overlay]]
*29/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/29|SDS-PAGE& Finalizing RII overlay]]
*30/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/30|Splitting cells, Immunoblot]]
*30/04/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/04/30|Splitting cells, Immunoblot]]
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<b>May</b>
*03/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/03|Split cells for RII overlay]]
*03/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/03|Split cells for RII overlay]]
*05/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/05|Put cells to S<sup>0</sup> for RII overlay
*05/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/05|Put cells to S<sup>0</sup> for RII overlay
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*28/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/28|Immunoblot AKAP79 (upstate)]]
*28/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/28|Immunoblot AKAP79 (upstate)]]
*31/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/31|SDS-PAGE for AKAP79 immunoblot & RII overlay (HEK293)]]
*31/05/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/05/31|SDS-PAGE for AKAP79 immunoblot & RII overlay (HEK293)]]
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<b>June</b>
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*01/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/01|SDS-PAGE for immunoblot (fail)]]
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*02/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/02|Preparing TRIS Urea gel]]
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*03/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/03|Tris urea gel]]
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*07/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/07|Urea-TRIS page, splitting cells]]
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*08/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/08|Creating RNA isolation protocol]]
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*09/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/09|Refreshed medium]]
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*10/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/10|Put cells to S<sup>0</sup>, split cells]]
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*11/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/11|Cell stimulation]]
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*12/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/12|Stop stimulation: Protein and RNA/Protein isolation]]
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*13/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/13|Putting cells to S<sup>0</sup>, splitting cells and continuing of protein isolation]]
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*14/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/14|Cell stimulation]]
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*15/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/15|Stop stimulation: Protein and RNA/Protein isolation (2)]]
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*16/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/16|Stimulation of cells]]
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*17/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/17|Stop stimulation: Protein and RNA/Protein isolation (3)]]
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*21/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/21|New HEK293, Bradford assay, cAMP precipitation]]
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*22/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/22|cAMP precipitation]]
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*23/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/23|Immunoblot]]
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*24/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/24|Immunoblot repeat]]
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*25/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/25|Coomassie staining]]
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*28/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/28|HeLa lysis, Bradford & cAMP precipitation part I]]
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*29/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/29|cAMP precipitation part II & SDS-PAGE]]
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*30/06/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/30|Lyse HeLa]]
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<b>July</b>
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*01/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/01|Bradford, qPCR AKAP12 & AKAP9, Silverstaining]]
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*02/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/02|qPCR GAPDH, Silverstaining]]
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*03/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/03|Silver staining attempt #3]]
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*05/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/05|Bradford & cAMP precipitation]]
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*06/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/06|cAMP precipitation gel]]
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*07/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/07|RII overlay cAMP precipation]]
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*08/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/08|RII overlay result]]
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*09/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/09|Gravin Co-IP, cAMP precipation gel]]
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*10/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/10|Gravin Co-IP gel]]
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*11/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/11|AKAP250 (scbt) immunoblot cAMP precipitation]]
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*12/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/12|AKAP250 (scbt) immunoblot AKAP250 Co-IP]]
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*{{todo}} 13/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/13|AKAP250 (Eurogentech) immunoblot]]
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*{{todo}} 14/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/14|AKAP250 (abcam) immunoblot]]
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*15/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/15|Epac immunoblot]]
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*{{todo}} 16/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/16|Epac immunoblot result]]
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*19/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/19|Epac immunoblot repeat, new cell culture]]
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*{{todo}} 20/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/20|Epac immunoblot repeat result]]
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*21/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/21|Splitting cells]]
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*24/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/24|Splitting cells for immunofluorescence]]
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*25/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/25|Cell check]]
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*26/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/26|Cover slips to S<sup>0</sup> & Plating out for ELISA]]
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*27/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/27|Stimulation cover slips, PKA-Co-IP]]
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*28/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/28|Immunofluorescence for cells, Gravin & RII localization]]
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*29/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/29|PKA-Co-IP gel]]
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*30/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/30|Trypsin/EDTA dilution, splitting cells for ELISA]]
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*31/07/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/07/31|Put cells to S<sup>0</sup>]]
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<b>August</b>
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*01/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/01|Stimulation for ELISA]]
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*02/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/02|Put cells to S<sup>0</sup>, Splitting cells, stopping stimulation]]
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*03/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/03|ELISA, RII, IF]]
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*04/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/04|Stop stimulation, put IF to S<sup>0</sup>]]
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*05/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/05|Start stimulation ELISA & IF, splitting cells]]
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*06/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/06|Ending stimulation ELISA, Primary antibody IF]]
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*07/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/07|Finish IF, Split cells]]
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*09/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/09|Isolate Protein/RNA Jurkat cells, Codes IF]]
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*10/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/10|ELISA, qPCR Jurkat, SDS-PAGE cAMP precipitation]]
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*11/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/11|Jurkat qPCR results, Primary antibody WB, Ending stimulation ELISA]]
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*12/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/12|Lyse HEK293 (isolate RNA), 2<sup>nd</sup> AB WB, Convocal microscopy]]
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*13/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/13|SDS-PAGE Lysates, strip blot]]
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*14/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/14|Secondary antibody cAMP precipitation blot of Gravin en AKAP79]]
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*16/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/16|SDS-PAGE lysates, Cell splitting]]
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*17/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/17|ON Blot lysate samples]]
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*18/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/18|BioRad workshop]]
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*19/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/19|Prepare cDNA, Immunoblot visualization, 1<sup>st</sup> AB ON Blot]]
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*20/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/20|Stripping blots, qPCR AKAP8, refresh medium new cells, 2<sup>nd AB]]
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*21/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/21|Lysate SDS-PAGE, qPCR AKAP5, Western blot AKAP79 and Gravin]]
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*23/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/23|SDS-PAGE cAMP precipitation, pre-IgG antibody]]
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*24/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/24|RII Overlay, Pre-IgG 2<sup>nd</sup> AB, CSE on HEK293, ASMC to S<sup>0</sup>, Split cells w. Mileh]]
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*25/08/2010: [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/08/25|RII Overlay all]]
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Project Description/Abstract

“Towards insights into cyclic AMP compartmentalization in cell of lung origin” a cooperation between the department of Molecular Pharmacology (University Groningen, the Netherlands) and the Leibniz-Institut für Molekulare Pharmakologie (Berlin, Germany). These are the entries of a Master internship from February 'till September 2010.

Notes

Interesting entries

Protocols

Results

ELISA
  1. Viability & ELISA
  2. Dose effect curve ELISA
  3. ELISA
  4. ELISA
  5. ..
  6. ELISA Berlin Okay
Co-IP
  1. Co-IP 1 Anouk
  2. Co-IP 2 Anouk
  3. Co-IP 3 Anouk
  4. Co-IP 4 TODO
RII Overlay
  1. ..
  2. RII overlay Okay
Misc.

Convenient Links

Downloads

Blanc for IL-8 ELISA (Linear regression)

Blanc for protein size on gel/membrane (non-Linear regression)

Blanc form IF coverslips

Table of contents

'
Groningen Berlin

February

March

April

May

June

July

August


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