Immunofluorescence Mountant - Revised 1/2/2006 MAS
SOLUTIONS and PROTOCOL:
10% Phosphate Buffered Airvol-205 (polyvinyl alcohol) with 2% n-propyl gallate.
Airvol-205 (Air Products and Chemicals Inc., Allentown, PA 18105)
5.0 g in 20.0 ml n-H2O
Disperse Airvol-205 into vortex of water in a 125 ml glass bottle. Allow to hydrate for 15 minutes.
Autoclave for 30 min. to solublize the Airvol-205. Cool to room temperature.
Dissolve 1.0 g n-propyl gallate (Sigma) in 25 ml glycerol (overnight on stir plate).
Mix n-propyl gallate and Airvol.
Add 0.35 ml 0.1M monobasic phosphate buffer.
Adjust pH > 8.0-8.5 with 0.1M dibasic phosphate buffer.
Q.S. with n-H2O if necessary to 50.0 ml.
1-2 µg/ml DAPI may be added to media to label DNA.
Aliquot and store at -20°C.
Note:
pH must be >8.0-8.5. Check solution before every use.
Centrifuge mountant before use.
Add small amount to slide (~15-20 µl per 22X50mm cover slip), add cover slip and press gently to remove excess mountant.
Place at 37°C for 10-15 minutes to polymerize Airvol. The cover slip can still be removed if necessary. Airvol may soften if slides is refrigerated. Cover slip may be sealed with nail polish to make a more permanent seal.
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