Adi spp 555

For in vitro protein-protein interaction experiments, we used biolayer interferometry technology (Octet Red, Forté-Bio, USA). Recombinant HSF2 (TP310751 Origen) was desalted (ZebaTM Spin Desalting Columns, 7 K molecular weight cutoff, 0.5 ml (1034–1164, Fisher Scientific, Germany)) and biotinylated at a molar ratio biotin/protein (3:1) for 30 min at room temperature (EZ-Link NHS-PEG4-Biotin [1189–1195, Fisher Scientific, Germany]). Excess Biotin was removed using ZebaTM Spin Desalting Columns. Biotinylated recombinant HSF2 was used as a ligand and immobilized at 100 nM on streptavidin biosensors after dilution in phosphate-buffered saline (PBS; 600 s). Interactions with desalted analytes diluted in PBS at 100 nM (recombinant CBP domains 6 His-tag Full-HAT, Bromodomain [BD], RING, or HSP70 as a positive control [ADI-SPP-555, Enzo-Life Sciences]) were analyzed after association (600 s). All sensorgrams were corrected for baseline drift by subtracting a control sensor exposed to running buffer only. For Kd determination, each Kd was calculated with a 1:1 stoichiometry model using a global fit with Rmax unlinked by sensor (FortéBio, Data analysis software version 7.1.0.89).

For in vitro protein-protein interaction experiments, we used biolayer interferometry technology (Octet Red, Forté-Bio, USA). Recombinant HSF2 (TP310751 Origen) was desalted (ZebaTM Spin Desalting Columns, 7 K molecular weight cutoff, 0.5 ml (1034–1164, Fisher Scientific, Germany)) and biotinylated at a molar ratio biotin/protein (3:1) for 30 min at room temperature (EZ-Link NHS-PEG4-Biotin [1189–1195, Fisher Scientific, Germany]). Excess Biotin was removed using ZebaTM Spin Desalting Columns. Biotinylated recombinant HSF2 was used as a ligand and immobilized at 100 nM on streptavidin biosensors after dilution in phosphate-buffered saline (PBS; 600 s). Interactions with desalted analytes diluted in PBS at 100 nM (recombinant CBP domains 6 His-tag Full-HAT, Bromodomain [BD], RING, or HSP70 as a positive control [ADI-SPP-555, Enzo-Life Sciences]) were analyzed after association (600 s). All sensorgrams were corrected for baseline drift by subtracting a control sensor exposed to running buffer only. For Kd determination, each Kd was calculated with a 1:1 stoichiometry model using a global fit with Rmax unlinked by sensor (FortéBio, Data analysis software version 7.1.0.89).

For in vitro protein-protein interaction experiments, we used biolayer interferometry technology (Octet Red, Forté-Bio, USA). Recombinant HSF2 (TP310751 Origen) was desalted (ZebaTM Spin Desalting Columns, 7 K molecular weight cutoff, 0.5 ml (1034–1164, Fisher Scientific, Germany)) and biotinylated at a molar ratio biotin/protein (3:1) for 30 min at room temperature (EZ-Link NHS-PEG4-Biotin [1189–1195, Fisher Scientific, Germany]). Excess Biotin was removed using ZebaTM Spin Desalting Columns. Biotinylated recombinant HSF2 was used as a ligand and immobilized at 100 nM on streptavidin biosensors after dilution in phosphate-buffered saline (PBS; 600 s). Interactions with desalted analytes diluted in PBS at 100 nM (recombinant CBP domains 6 His-tag Full-HAT, Bromodomain [BD], RING, or HSP70 as a positive control [ADI-SPP-555, Enzo-Life Sciences]) were analyzed after association (600 s). All sensorgrams were corrected for baseline drift by subtracting a control sensor exposed to running buffer only. For Kd determination, each Kd was calculated with a 1:1 stoichiometry model using a global fit with Rmax unlinked by sensor (FortéBio, Data analysis software version 7.1.0.89).