User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/06/29

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] cAMP precipitation part II & SDS-PAGE
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Washing beads: cAMP precipitation

 * Spin beads 5 min. 2100xg @ 4 °C
 * Collect supernatant (= NAC)
 * Add 500 μL Potter buffer
 * Spin 5 min. 2100xg @ 4 °C
 * Remove supernatant with vacuum pump
 * Add 500 μL Potter buffer
 * Spin 5 min. 2100xg @ 4 °C
 * Remove supernatant with vacuum pump
 * Add 500 μL Potter buffer
 * Spin 5 min. 2100xg @ 4 °C
 * Remove supernatant with vacuum pump and syringe
 * Add 50 μL of 4x sample buffer
 * Spin 5 min. 2100xg @ 4 °C

Protein work

 * SDS-PAGE
 * RII Overlay
 * Silver / Coomassie staining
 * Immunoblot
 * AKAP250
 * AKAP95
 * AKAP450

RNA work

 * Preparing cDNA
 * Real time qPCR