Bind Silane and 48 well combs - the answers
Phillip San Miguel
pmiguel at bilbo.bio.purdue.edu
Sun Feb 1 10:57:28 EST 1998
We use bind silane with Alf express gels. I hate the stuff, but it's a
necessary evil if you want to keep the sides of the wells from moving around.
The recipe pharmacia gave us uses too much bind silane by more than a factor of
ten. We treat both plates but only in the area where the wells will be. Every
four or five months I add 3.5 ul of Bind Silane to 10 ml of Ethanol. I keep
this as a stock solution at room temp. It doesn't seem to go bad. Just before
assembling plates, I add 60 ul of the stock solution and 15 ul of 10% acetic
acid to a microfuge tube and mix. I use about 10 ul on a plate -- drawing a
thin bead of liquid across with a pipettor then wiping it into an area about 1
cm wide with a kim-wipe. Then I squirt ddH2O on the area and polish and do the
same with isopropanol.
After running the gel and taking the gel plates apart a paper towel is
pressed on the gel and removed. The acrylamide only sticks to glass in the
area where the wells are. The rest goes with the paper towel. We have a
plastic scraper to remove the acrylamide left at the top of the gel. I don't
worry much about scratching the top edge of an Alfx gel -- the detectors are
near the bottom. Then we wash as normal, using a teflon safe scrub sponge to
remove the residual acrylamide from the top of the gel. I don't use it on the
rest of the glass plate -- it will scratch the glass on occasion. The
cellulose side of the sponge will clean the rest of the plate if you didn't use
a huge amount of bind silane at the top.
I think too much bind silane can cause problems other than getting the
plates clean. A Pharmacia rep said that an "ion front" migrates in the gel
from the bind silane. He suggested that fluctuations we were seeing in our
voltage during a run were caused by this.
Phillip San Miguel
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