bsh at MED.PITT.EDU
Thu May 26 17:12:58 EST 1994
> I have used Sigma A4503 many times to make Denhardt. I
> usually make 200X Denhardts. Indeed, it is slow to
> dissolve, but with a stirring bar and time it works.
> Also it is helpful to apply some heat --not too high
> or you could get denaturation , but 40-45 deg C.
> Good luck.
> rocket1 at bu.edu
One of the common mistakes I have seen made in trying to
dissolve BSA, among other things, is that we try to take the BSA
powder in a beaker, add the required amount of DW and start shaking.
When we do this, if you see closely, the BSA powder forms a gluey
lump immediately on coming contact with water. And it is this
lump that takes a long time for dissolving.
Instead, I usually have the beaker with water on a magnetic
stirrer and stirring. Now, when I add the BSA powder, it does not
form a lump, but instead will form something similar to a suspension.
Stirring for few minutes will make this go into the solution
as opposed to the long time it takes for a big lump. If anybody
has some experience making a solution of caesin for blocking western
blots and trying to adjust the pH with acid, you may recall that
the same holds true.
Just my 0.000001c worth.
bsh at med.pitt.edu
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