User:Karmella Haynes/Notebook/PcTF Genomics/2015/08/14

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08/14/15

  • Western Castone - washes, secondary staining, imaging

Western Castone - washes, secondary staining, imaging

  • Washed 3x 10 min. in PBST
  • Used "floating" technique to stain w/ 500 μL 1:1000 goat anti-rabbit-HRP secondary in 5% milk/PBST
  • Stained for 1 hour/ room temp
  • Washed 3x 10 min. in PBST
  • Used Western Femto from Thermo fisher; inverted membranes onto 500 μL working solution; 5 min./ room temp


RESULTS

  • Ladder: Pageruler Plus: [1]
  • dCas (no insert) = 166.2 kD (predicted)
  • Histones = ~15 kD


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