hello everyone:
recently I found two proteins interaction in the gel filtration. the result can be repeated and the best ratio for mixture is 1:1. but the negative results always when we do the GST Pull-down to trail this interaction. the detail : we use 10mg/ml for every protein and mix together for 2h on the ice. then commix with the GST-beads for 30 min. 12000rpm in centrifuge.discard the suspending and wash the beads with 1x PBS three times. SDS-PAGE found no interaction between targets.
please give me your idea.
Sudan
