My primary antibody contains many additives: what should I do?

For pre-treatment of the antibody before using Duolink Probemaker, we refer to standard procedures.

To change buffer and/or remove low molecular additives like azide or Trehalose you can either dialyze against PBS or perform gel filtration on a spin column of e.g. Sephadex G25 or G50 (GE Healthcare) equilibrated with PBS.

To remove high molecular additives like BSA or gelatin we recommend purification of protein G or A columns (available from e.g. GE Healthcare). The antibody is eluted with low pH which might affect antibody activity and we therefore recommend compensating with the immediate addition of strong buffer of neutral pH. There is always a loss of antibody amount in this procedure which could be quite substantial.