User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/06/01

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SDS-PAGE for immunoblot (fail) Main project page
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Summary

  • Gel #2 was tried with fresh APS 10% (previous was 1 month old)
    • Still large blur was seen during and after running the gel.
    • When checking pH of the stacking gel a pH of 7.15 was found instead of 6.8
    • This will prevent the stacking function and would explain the blur seen while running
  • New stacking buffer was prepared
    • 18.9 g TRIS
    • 150 mL dH2O
    • pH 6.8
    • added to 250 mL with dH2O
  • Gel #3 was prepared
    • All other gels stored at 4 °C were discarded
    • To prevent more wasting of samples only marker and loading buffer (4x) was used
    • seemed to run owkay after also running buffer was renewed
  • Immunotreatment of blot #2 was performed as described yesterday

Materials & Methods

Materials

Method

#2 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
SamplehTERT RIPAhTERT PotterHEK293 RIPAMarkerhTERT RIPAhTERT PotterHEK293 RIPAMarkerhTERT RIPAhTERT PotterHEK293 RIPAMarkerhTERT RIPAhTERT PotterHEK293 RIPA


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