B/B Washout Ligand

Description

B/B Washout Ligand is a membrane-permeant ligand that dissociates protein interactions induced by the B/B Homodimerizer, which is provided in each of the iDimerize Inducible Homodimer Systems Cat. Nos. 635068, 635072, 635084, and 635086 . The B/B Washout Ligand dissociates these interactions with aT1/2of 10 minutes after adding it to target cells treated with B/B Homodimerizer. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back B/B Washout Ligand releases B/B Homodimerizer-induced interactions.HeLa cells were transfected with a bidirectional vector Panel A which expressed two fluorescent proteins tdTomato and AcGFP1 each fused to a DmrB dimerization domain. The tdTomato protein was additionally tagged with the FLAG epitope DYKDDDDK to enable efficient immunoprecipitation. After overnight treatment with 0.1 muM B/B Homodimerizer, transfected cells were lysed and immunoprecipitated using anti-DYKDDDDK beads Cat. No. 635686 which recognize the FLAG epitope Panel B . The beads were washed three times with buffer containing both 0.1 muM B/B Homodimerizer and increasing concentrations of B/B Washout Ligand. After washing, the beads were boiled in SDS-sample buffer, centrifuged, and separated on a polyacrylamide gel. The samples were then analyzed via Western blot using an anti-AcGFP1 specific antibody, and an anti-FLAG antibody to detect the tdTomato fusion Panel C . B/B Washout Ligand efficiently competed for binding to the DmrB tagged proteins, releasing DmrB-AcGFP1 from the beads. Back Back