User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/07/08: Difference between revisions

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**1% Igepal CA-630
**1% Igepal CA-630
**1% deoxycholic acid
**1% deoxycholic acid
**pH 7.4
*Addition
**1 mM NaF
**1 mM NaF
**1 mM Na<sub>3</sub>VO<sub>4</sub>
**1 mM Na<sub>3</sub>VO<sub>4</sub>
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**7 µg/ml pepstatin A
**7 µg/ml pepstatin A
**1 mM PMSF
**1 mM PMSF
**pH 7.4
 
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==Related==
==Related==

Revision as of 07:12, 8 July 2010

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Summary

  • * RII overlay, washing of membranes and taking pictures
    • Other membranes looked okay, for my membrane see below

Results

  • RII Overlay 3h autoradiograph

  • Only RII subunits seem to be visible, HEK293 has a band around 66 kDa which should at least be >75 kDa (AKAP79)

Discussion

  • Maybe a new lysis buffer should be better?
  • RIPA BUFFER GRONINGEN
    • 40 nM Tris
    • 150 mM NaCl
    • 1% Igepal CA-630
    • 1% deoxycholic acid
    • pH 7.4
  • Addition
    • 1 mM NaF
    • 1 mM Na3VO4
    • 10 µg/ml aprotinin
    • 7 µg/ml leupeptin
    • 7 µg/ml pepstatin A
    • 1 mM PMSF


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