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)
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