DSS (disuccinimidyl suberate) crosslinking in situ

This protocol describes how to use DSS to crosslink primary amines in situ.

Figure. DSS. Disuccinimidyl suberate is a six-carbon lysine-reactive non-cleavable cross-linking agent. It consists of functional groups It is a homobifunctional N-hydroxysuccinimide ester formed by carbodiimide-activation of carboxylate molecules, with identical reactive groups at either end. Because of its powder form it has longer shelf-life than pegylated NHS crosslinkers like BSPEG.


1️⃣Begin by measuring out DSS powder and dissolve in DMSO and then add the pH 8 buffer

Reagent Amount Final concentration
DSS 2 mg 12.5 mM
Anhydrous DMSO 432 µl
1xPBS pH 8 277 µl

2️⃣ Add to the sample and incubate for 1h at room temperature.

3️⃣Quench the reaction by aspirating and adding 1 M Tris pH 8.0 and incubate for 30 min.

4️⃣Wash.

5️⃣The sample is now ready.