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

 .todo { color: red } .done { color: green} {| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Ht31P toxicity
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

Summary

 * Stop stimulation
 * Prepare washing buffer 1x ELISA + 10x PBS
 * CoIP

Preparations for ELISA & Viability staining

 * As described 17Feb2010

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



 * Viability staining



Conclusion

 * Ht31P is lethal for airway smooth muscle hTERT cells
 * Strength of effect is donor dependent


 * }