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

From OpenWetWare
Jump to navigationJump to search

<html><style type="text/css"> .todo { color: red } .done { color: green} </style></html>

SDS-PAGE for immunoblot (fail) <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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
Sample hTERT RIPA hTERT Potter HEK293 RIPA Marker hTERT RIPA hTERT Potter HEK293 RIPA Marker hTERT RIPA hTERT Potter HEK293 RIPA Marker hTERT RIPA hTERT Potter HEK293 RIPA


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/06/01&oldid=446319"