User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/04/07

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Preparation first Pilot Berlin
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Summary

 * Stimulated cells with CSE 15% @ 12 o'clock

Materials

 * Warm DMEM S0
 * Warm Dulbecco’s phosphate buffered saline (PBS)]

Induction 10cm dishes: cAMP precipitation and RII overlay

 * Prepare CSE 100% (25 mL) (DMEM S0)
 * Prepare CSE 15% (6 mL 100% in 40 mL with DMEM S0)
 * Wash twice the (3) 10cm dishes with PBS
 * Add 10 mL of CSE 15% to 2 of 3 plates and S0 to the third
 * Incubate 24 h @ 37 °C

Prepare cells from -80 °C

 * Cells were plated out from the -80 °C freezer as described 23March2010
 * D9 P21 (prepped 22Jan2010)
 * D12 P26

Prepare DMEM S+

 * FCS was heat inactivated as described 23March2010
 * All 50 mL was put into DMEM, together with 10 mL PenStrep

Related topics

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