Senescent-associated b-Gal Assay
Wash cells with PBS Fix cells in 0.2% glutaraldehyde for 5’ dilute this in PBS W
Wash cells with PBS Fix cells in 0.2% glutaraldehyde for 5’ dilute this in PBS Wash cell with PBS Stain with X-gal solution (ingredients listed below) for 6 to 24 hours at 37 °C
Chemical Conc. | Stocks | Amount needed for 10 plates (40 ml) |
| 1 mg/ml X-gal | 40 mg/ml | 1 ml |
| 150 mM NaCl | 5 M | 1.2 ml |
| 2 mM MgCl2 | 1M | 80 μ l |
| 5 mM K3 Fe(CN)6 | 100 mM | 2 ml |
| 5 mM K4 Fe(CN)6 | 100 mM | 2 ml |
| 40 mM NaPi, pH 6.0 | 0.1M | 16 ml 17.72 ml dd H2 O |
NaPi - 0.1M at pH 6.0
- 2.5 ml Na2 HPO4
- 17.6 ml NaH2 PO4
- dd H2 O to 200 ml
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