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(Janine’s protocol)
1. Add 200µl of acetone, vortex briefly.
2. Remove acetone and pipette into flat-bottom 96-well plate (polystyrene)
3. Centrifuge plate at 3000 x g for 3-5 minutes.
4. Invert plate carefully and blot dry with paper towels
5. Add 40µl of X-Gluc stain solution to each well (multi-pipettor) and incubate at 37°C overnight.
X-Gluc Solution 1 20 mg/ml X-Gluc concentration. e.g. For a 1.5 ml tube (microfuge tube) dissolve 30 mg (.03 g) of X-Gluc in 1.5 ml DMSO (dimethyl sulfoxide).
2 Cover with foil.
3 Vortex well.
4 Keep at -20°C.
X-Gluc stain solution
*Note: 4ml of stain solution are necessary to stain 1 96-well plate.
| Stock solution (10x)
50 mM ferrocyanide
50 mM ferricyanide
500mM NaPO4, pH 7
20mg/ml X-Gluc (in DMSO)
ddH2O |
Working solution (1ml)
100µl
100µl
25µl
775µl
1000µl = 1 ml |
- Cover with foil
- Keep at 4°C.
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