ß-gal Staining:
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Reagents: |
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0.1M Phosphate Buffer (pH 7.3): |
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0.1M Sodium Phosphate monobasic |
115ml |
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0.1M sodium phosphate dibasic |
385ml |
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Total Volume |
500 ml |
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Fix solution: |
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0.5% Gluteraldehyde: |
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25% gluteraldehyde |
0.4 ml |
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100mM EGTA pH 7.3 |
2.5 ml |
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1M magnesium chloride |
0.1 ml |
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0.1 M sodium phosphate pH 7.3 |
47.0 ml |
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Total Volume |
50.0 ml |
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4% Paraformadehyde: |
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paraformadehyde |
20 g |
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1M magnesium chloride |
1ml |
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100mM EGTA |
25ml |
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~500 ml PBS |
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Wash buffer: |
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1M magnesium chloride |
0.4 ml |
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1% deoxycholate |
2.0 ml |
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2% Nonidet-P40 |
2.0 ml |
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0.1M sodium phosphate pH 7.3 |
195.6 ml |
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Total Volume |
200.0 ml |
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X-gal Staining: |
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25mg/ml x-gal stock dissolved in di-methyl formamide |
2.0 ml |
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potassium ferrocyanide (Sigma P-9387) |
0.106 g |
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potassium ferricyanide (Sigma P-8131) |
0.082 g |
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wash buffer |
48.0 ml |
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Total Volume |
50 ml |
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- This buffer can be reused, filter after use and store in the dark.
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- Also note, crystal from due to di-methyl formamide. If these crystal
are a problem, prepare X-gal stock in dimethyl sulfoxide. |
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