User:Karmella Haynes/Notebook/PcTF Genomics/2015/02/03

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02/03/15

  • Cytology of KAH126 cells - blocking

Cytology

  • 2/02/15: Fix & permeabilize
    • Wash 1x w/ R.T. PBS
    • Fix: 4% formaldehyde/PBS
    • Permabilization: 5% Triton/PBS (R.T./ 10 min.)
    • Wash 1x w/ R.T. PBS
    • Store in 1x PBS overnight at 4°C

02/03/15
> Block: Immuno Protein Blocking Solution with Horse Serum (Gentex GTX30974), 4°C/ overnight.


TOMORROW...

  • Primary stain (rabbit polyclonal)
    • "Face down" method (see 3/30/10); 20 μL diluted ab; R.T./2.5 hrs.
      • H3K27me3 07-449, 1:200 in Immuno Protein Blocking Solution with Horse Serum (Gentex GTX30974)
  • Wash w/ 1x PBS, 4x/R.T./5 min.


  • Secondary stain
    • "Face down" method (see 3/30/10); 25 μL diluted ab; R.T./1 hr.
      • goat anti-rabbit-Alexafluor488, 1:1000; Hoescht, 1:1000
  • Wash w/ 1x PBS, 4x/R.T./5 min.


  • Mount samples w/ Vinol solution; allow to set > overnight (in dark)