Recipes
Phosphate-buffered saline
| Salt |
Conc. |
Mol. Wt. |
Grams/Liter |
| NaCl |
137 mM |
58.44 |
8.0 |
| KCl |
2.7 mM |
74.56 |
0.2 |
| KH2PO4 |
1.5 mM |
136.09 |
0.2 |
| EITHER: Na2HPO4 |
8.0 mM |
141.96 |
1.14 |
| OR: Na2HPO4·7H2O |
8.0 mM |
268.07 |
2.14 |
| • Adjust pH to 7.2 with 2 N HCl (should take 10-12 drops for 1 liter). |
| • Add sodium azide to 0.01%. Omit azide if PBS is to be used with live tissue. |
| • Our PBT is 0.1% Triton X-100 (1 ml/l). |
| • For antibody staining we use PBT + 0.1% BSA. |
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Poly-L-lysine coated coverslips or slides
| We use Sigma #P-1524. |
| polylysine, 0.06% |
6 mg |
12 mg |
36 mg |
100 mg |
| Kodak PhotoFlo |
20 ul |
40 ul |
120 ul |
330 ul |
| RO water |
10 ml |
20 ml |
60 ml |
165 ml |
| • Add sodium azide if desired to 0.01%; keep refrigerated. |
| • Dip slides or coverslips briefly, air dry, rinse in distilled water, air dry again. Use within 2-3 days. |
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Murray Clear series
Murray Clear is 2 parts Benzyl Benzoate to 1 part Benzyl Alcohol (by volume). This should have a refractive index of about 1.55.
For compatibility with phalloidin, use an Isopropanol series to transition from PBS to Murray Clear: 1 x 70%, 1 x 85%, 1 x 95%, 2 x 100%, followed by 3 x Murray Clear. Otherwise use 3 x 100% Methanol
Calcium/Magnesium-free artificial seawater
| Stock |
50 ml 2x |
100 ml 2x |
200 ml 2x |
| 2 M NaCl |
22.5 ml |
45 ml |
90 ml |
| 0.33 M Na2SO4 |
10 ml |
20 ml |
40 ml |
| 0.5 M KCl |
1.8 ml |
3.6 ml |
7.2 ml |
| 0.5 M NaHCO3 |
0.5 ml |
1.0 ml |
2.0 ml |
| 0.2 M Tris pH 7.8* |
5 ml |
10 ml |
20 ml |
| 0.5 M EDTA |
0.2 ml |
0.4 ml |
0.8 ml |
| RO water |
10 ml |
20 ml |
40 ml |
| • Make up separate stock solutions in plastic containers, and store refrigerated. Do NOT add sodium azide. |
| • * The pH of Tris solutions is sensitive to dilution and temperature; check pH of stock solution at the final concentration and temperature, and adjust until it is pH 8±0.2 |
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