User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/02/24

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Stimulation hTERT cells v2 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Summary

Conditions

  • CTR
  • CSE
  • CSE + 8-pcpt
  • CSE + 6-Bzn
  • 8-pcpt
  • 6-Bnz
  • CSE + Rp-8-CPT-cAMPS
  • Rp-8-CPT-cAMPS
  • CSE + siRNA
  • siRNA

Materials & Methods

Materials

Method

Co-immunoprecipitation

  • Use cells put on S0 23Feb2010
  • Add ... μL Lysis buffer
  • Spin down beads prepared 23Feb2010
    • Remove liquid
  • Add PBS to wash
  • Spin down and remove liquid
  • Prepare
  1. Only beads (100 μL)
  2. Beads w. antibody (100 μL)
  3. Beads w. antibody (100 μL) & lysis buffer (200 μL)
  4. Beads w. antibody (100 μL) & sample (200 μL)

(possible also beads (100 μL) with only sample (200 μL))

  • Incubate?
  • Wash?
  • Add 75 μL 4x loading buffer
  • Boil samples and put on gel


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