Anti gst antibody

Purified GST‐tagged and His₆‐tagged proteins were used, respectively, as bait and prey proteins. 100 μL 50% glutathione‐sepharose beads (equilibrated with PBS buffer) and GST‐tagged protein (final concentration 5 nM) were incubated for 2 h at 4°C, and the mixture was then added with His₆‐tagged protein (final concentration 5 nM) and incubated for another 2 h at 4°C. A mixture of GST and His₆‐tagged prey protein was used as negative control. After centrifugation, beads were washed 5× with PBS buffer, and the resulting pellet was added with 100 μL elution buffer (PBS buffer containing 10 mM glutathione) to release protein from beads. The mixture was centrifuged, and supernatant containing bait and prey proteins was collected and analysed by SDS‐PAGE. Separated proteins were identified by Western blotting using anti‐His₆ (Sigma) or anti‐GST antibody (Cwbio, China).