User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/03/17: Difference between revisions

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===Method===
===Method===
====Co-immunoprecipitation====
====Preparations for ELISA & Viability staining====
* As described [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/17| 17Feb2010]]
====[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/25|Co-immunoprecipitation Part III]]====
* Centrifuge sample incubated with beads, 5 min. 14.000 RPM
* Centrifuge sample incubated with beads, 5 min. 14.000 RPM
* Remove supernatant<sup>*</sup> (w. syringe)
* Remove supernatant<sup>*</sup> (w. syringe)
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* Add 75 μL Laemli buffer to beads
* Add 75 μL Laemli buffer to beads
* Store samples @ -20 °C
* Store samples @ -20 °C
** Alternatively go directly on to Western blot (Boil samples and spin beads down, bring supernatant onto gel)
 




<sup>*</sup> Be careful not to touch beads
<sup>*</sup> Be careful not to touch beads
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==Results==
==Results==

Revision as of 06:09, 17 March 2010

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

Project name <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

  • Stop stimulation
  • Prepare washing buffer 1x ELISA
  • CoIP

Materials & Methods

Materials

Method

Preparations for ELISA & Viability staining

Co-immunoprecipitation Part III

  • Centrifuge sample incubated with beads, 5 min. 14.000 RPM
  • Remove supernatant* (w. syringe)
  • Add 1 mL PBS
  • Centrifuge, 5 min. 14.000 RPM
  • Remove supernatant*
  • Add 75 μL Laemli buffer to beads
  • Store samples @ -20 °C


* Be careful not to touch beads

Results

  • WHAT?

Conclusion

  • DONE Ow..



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