Isopeptidase T

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Biochemist986's picture
Isopeptidase T

i have a protien that i think may be ubiquitinated. after treating my cells with a proteasome inhibitor i see a band with slower mobility on a western blot leading me to believe it is ubiquitinated. One thing i want to try is using the deuquitinating enzyme Isopeptidase T. The idea is i treat my cell lysates with isopeptidase t, and if my protein is ubiquitinated the slower band i see on my western should go away. I was wondering has anyone ever done this and is there any Lysis buffer you can recommend ??