User:Karmella Haynes/Notebook/PcTF Genomics/2013/06/13

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06/13/13

  • Gal4-EED/luc cells IFC - secondary stain & imaging



Gal4-EED/luc cells IFC

  • Samples: first row of wells in the 24-well plate.
  • Keep all others under PBS
  1. 0 μg/mL dox
  2. 0.1
  3. 0.2
  4. 0.5
  5. 1.0
  6. 2.0
  • Follow standard protocol; wash 3x with 1xPBS
  • Use 1:1000 dilution of anti-rabbit Alexa488 and 1:1000 Hoescht (in 15 μL Gentex horse serum blocking solution per well)
  • Incubate under parafilm "covers" for one hour at room temperature in light-tight box with wet paper towels
  • Use GFP setting on Nikon scope to record images

Results:

  • Saw some cells with very bright signal, some dim in 0 μg/mL dox well
  • Signal appeared to be dimmer overall for all others.




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