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Rhodamine-Phalloidin/Calcofluor Staining
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Reagents
- Formaldehyde - Polysciences, 10% EM Grade Cat #04018
- 10XPBS - 80 grams NaCl + 2 grams KCl + 14.4 grams Na2HPO4 + 2.4 grams KH2PO4. pH upon dilution to 1X should be 7.2.
- Rhodamine Phalloidin - Molecular Probes Inc., Dissolved in methanol according to manufacturers instructions to ~6.6uM and stored dessicated at -20¡C. Cat#R-415.
- Calcofluor - aka fluorescent brightener 28. Sigma Cat#F-6259. Dissolve to 1mg/ml in H2O overnight and store at 4¡C in the dark.
- Mount - Dissolve 100 mg p-phenylenediamine in 10 mls PBS, adjust the pH to above 8.0 with 0.5 M Na Carbonate buffer (pH 9.0) and bring the volume to 100 mls with glycerol. Add Dapi to 50 ng/ml. Mix thoroughly and store at -20¡C. It turns brown when it is bad.
