I would like to use 50 mM DTT in my buffer for gel filtration, but I can't find if it's compatible with the Superdex 200 10/300 GL (17-5175-01) column. Has anyone tried it yet?
Since Superdex 200 represents cross-linked agarose and dextran you don't have to worry about its compatibility with DTT. There should be no problem to use DTT in your buffer. Just remember that DTT is not very stable in solution (half-life 40 h at pH 6.5 and 1.5 h at pH 8.5). So, your buffers should be fresh prepared every time. Also, your proteins under reduced conditions might show their molecular weight higher than in their native non-reduced conformation after separation on the column..
Since Superdex 200 represents cross-linked agarose and dextran you don't have to worry about its compatibility with DTT. There should be no problem to use DTT in your buffer. Just remember that DTT is not very stable in solution (half-life 40 h at pH 6.5 and 1.5 h at pH 8.5). So, your buffers should be fresh prepared every time. Also, your proteins under reduced conditions might show their molecular weight higher than in their native non-reduced conformation after separation on the column..
Since Superdex 200 represents cross-linked agarose and dextran you don't have to worry about its compatibility with DTT. There should be no problem to use DTT in your buffer. Just remember that DTT is not very stable in solution (half-life 40 h at pH 6.5 and 1.5 h at pH 8.5). So, your buffers should be fresh prepared every time. Also, your proteins under reduced conditions might show their molecular weight higher than in their native non-reduced conformation after separation on the column..
Hi there,
DTT can be used on Superdex200. But 50mM is a much higher concentration than the usual one for DTT in SEC (1-5mM)...
Thank you both for your answers.
I also got the reply from the manufacturer's technical services and they say it should be compatible with up to 10-20 mM DTT and most likely up to 50 mM DTT.
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