SPombe3

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!)

HELP!!

One of the students in our lab accidentally resuspended our 200ug Anti-GFP antibody in 500uL of 20% SDS solution instead of sterile water!! (The bottles were right next to each other)

What should I do from here??
As the lab manager, I am responsible for all of the reagents, and antibodies are way too expensive to just order more!
Since antibodies are proteins, is our stock completely ruined now??


I tried to think of some steps I could take from here:
1. Use them as is (will the antibodies even work now?)
2. Use lyophilization to dry out the samples in our vacuum/centrifuge. However, I'm afraid the SDS will precipitate with the antibody.

Any other ideas??