Hello,

I have been experiencing problems with mowiol preparation for immunoflorescence staining. The problem is that the mounting media i get is of very high viscosity, the aliquots that i stored in -20 degrees are already solidified.

This is the procedure on how prepared my mowiol. Kindly, advise me on this.

Procedure

1. Put 6g glycerol in a 50ml plastic centrifuge tube and add a small stir bar.

2. Add 2.4g Mowiol (CAS number: 9002-89-5); stir to mix.

3. While stirring, add 6ml distilled water and leave for 2 hours at room temperature.

4. Add 12ml 0.2 M Tris (ph 8.5).

5. Incubate the tube in hot water (50°C) for 10 minutes to dissolve solids. This can be repeated, if necessary.

6. Centrifuge at 5000 xg for 15 minutes to remove any undissolved solids, using 50ml centrifuge tube.

7. Store 1ml aliquots in eppendorf 1.5 ml tubes at -20°C.

I will appreciate for your advises on this. thanks