Hello,
I need some tips on how to make a 6X SDS loading buffer for use in SDS PAGE.
My biggest problem is getting the SDS into solution.
Last time I made it, I heated the solution on the lowest possible setting on our stir/heating plate to get it into solution. Once the SDS appeared to be dissolved, I turned the heat off and added the DTT and bromophenol blue. While I was aliquotting the solution for storage, it almost seemed to be becoming more solid. Indeed, I just removed an aliquot from our -20C and it is impossible to pipette and seems to be more of a gel-like solid than a liquid. (Our 6X loading buffer has always been thick in the past, but not this thick). I also have tried making the buffer without heating at all, and the solution becomes super thick even quicker.
This is the recipe we use for 10mL:
3.75mL 1M Tris pH 6.8
1.2g SDS (solid)
6mL glycerol (100% stock)
0.006g bromophenol blue
0.462g DTT
Any help would be greatly appreciated, or if anyone has a protocol for making a 6X SDS loading buffer that works for them I would also appreciate that! (Please tell me how you get the SDS to dissolve without turning the solution into a solid!)
