Acid Washing Beads Peter Novick Lab, Department of Cell Biology Yale University School of Medicine http://info.med.yale.edu/cellbio/Novick/Second/Protocols/AcidBeads.pdf Materials:
1) 0.5 mm glass beads (Biospec Products) 1-4 lbs
2) 1% Triton-X100
3) 95% EtOH
4) 6N HCL (Conc. is 12N)
5) 6N HNO3 (Conc. is 15N)
6) Large glass beaker (at least 4x the volume of glass beads)
7) Glass Stirring rod
Procedure:
1) First make sure there is plenty of water in the distillation tank
and make sure the deionized water feed is running. This washing
goes through an enormous amount of distilled water so make sure
there will be some left for others to use when your done!
2) Add the beads to the beaker and fill the beaker with ddH20. Add
TritonX-100 to 1% and stir with the glass rod until the detergent is
dissolved. Continue to stir intermitently for 10 more minutes.
Carefully pour off the fluid and repeat the wash for another 10 min.
Rinse 2X with ddH20. Note: for each of the following washes you
should stir for at least 2 minutes before pouring off.
3) Wash 3-5 times with 95% EtOH (use the cheap stuff in the red
can). Rinse 2X in ddH20.
4) Wash 2X with 6N HCl. Obviously you need to be careful with all
the acid solutions in this procedure, wear gloves etc. Rinse once
with ddH20.
5) Wash 2X with 6N HNO3.
7) Rinse 3X in ddH20, then let it sit 5 min with stirring, repeat
twice and then until the pH of the rinse is aproximately the same as
the ddH20 from the tap (usually ~pH 6). You may want to rinse once
with 10 mM Tris pH 7-8 to speed things up (but make sure the Tris is
washed away before baking)
8) Drain as much of the fluid as possible from the beads and let
them air-dry overnight. Cover the beaker with foil and bake at
350°C for 2-3 hrs. Store in the cold room.
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