Recommended Anti-Fade for fluorescence microscopy – especially good for EGFP and other low expression reporting molecules.
Mowiol goes into solution with difficulty. It’s best to make a large batch and freeze aliquots at -20ºC.
Takes about 8 hours to prepare correctly
Prepare in clean 250 ml flask or beaker
- 24g glycerol (analytical grade)
- Add 9.6g Mowiol 4-88 (Fluka, #81381 – through Sigma) and stir thoroughly to mix
- Add 24ml UHP distilled water and leave for 2hr at room temperature
- Add 48ml 0.2M Tris-HCl buffer, pH8.5 **final volume will be 200 mls
- Stir with a clean stir bar on a hot plate on warm – heat gently to 50ºC stirring continually – at least 4-5 hours until the majority of the Mowiol powder goes into solution
- Aliquot into 50 ml centrifuge tubes, weigh and balance
- Centrifuge at 5000g for 15 min., there will be a pellet at the base, carefully remove supernatant
- Aliquot into 15 ml conical tubes – add only 10 mls for expansion
- Store at -20ºC for 12 months – store at room temperature no more than one month
Note for stereology: if you have sections mounted in DPX – remove the DPX by soaking and washing in zylene, rehydrate in PBS and Azide (for 1-2 nights) then mount in Mowiol