Dialysis is a relatively slow process, on the time scale of hours. If you want to remove the DTT and change the buffer quickly, use a desalting spin column. Examples of commercial ones include PD-10 (Cytiva), Zeba (ThermoFisher) and Micro Biospin (Bio-Rad). These take only a few minutes. They are appropriate for use with proteins or other macromolecules.

If the volume of your sample is not too high, i suggest you to perform desalting instead dialisys with is quite fast

Small volumes (up to 2-2,5ml) can changed of buffer using gravity flow pd-10 coloumns (see a video tutorial at: 8https://www.blogger.com/video.g?token=AD6v5dwNlVHz3BclDT9RLY-6KbEEqkS3RnRdHn4BqTsZW4j_prf3KZwHDMTzRYT-B9Yifvmde7RdExL5AArLFtg1Q13fu4-83o3M8bCka1jJ5PfWFK8B53BOpx6YrzvLr2qDiPgT15c)

while larger volumes (e.g 15ml) can be exchanged using a prepacked Hiprep26/60 coloumn connect to an chromatograpic sistem. It is a low pressure coloumn (0.5MPa maximun pressure) that may work well also in AKTA prime, AKTA start, AKTA Go and other simple chormatograpic sistems.

best regards

Manuele

Adam B Shapiro Manuele Martinelli Thank you for the suggestions. I have a 10 mL sample volume and don't have the equipment to connect to a chromatographic system. I have PD-10 desalting columns and was planning to use the spin protocol instead of gravity. However, according to the protocol I can only put a sample volume of 2.5 mL. I was wondering if I can keep adding multiple batches of 2.5 mL sample to the same PD-10 column after sample separation or use multiple PD-10 columns. Hope they are reusable.

PD-10 columns are reusable, in principle, but you would have to wash out the DTT before the next use. That would take a lot of washing to be sure to get rid of all of it.* Also, I'm not sure how well the resin would stand up to multiple uses as a spin column. Use 4 columns, if you can afford to. If you use them in drip mode, you can wash out the DTT at your convenience and be confident that they will be reusable.

*An easy test for DTT in the eluent is to use Ellman's reagent (5,5-dithio-bis-(2-nitrobenzoic acid)), which develops a yellow color (measure absorbance at 412 nm) in the presence of sulfhydryl groups.