A study published in GenomeBiology discusses a novel and advanced protein degradation technology that provides functional insights into how CTCF protein regulates DNA transcription.
cells were treated with DMSO or 10 μM 5-Ph-IAA for 6 h. Cells were fixed with 2% formaldehyde for 10 min at room temperature. Fixed pellets were lysed in 500 μL of ice-cold HiC lysis buffer with PI at 4 °C with rotation for 20 min followed by centrifugation at 2500 × g for 5 min at 4 °C. The pelleted nuclei were washed once with 500 μL of ice-cold HiC lysis buffer + PI, resuspended in 100 μL of 0.5% SDS, and incubated at 62 °C for 10 min with no shaking.
antibody [8 μg Diagenode rabbit polyclonal antibody ] was added and incubated at 4 °C overnight with rotation. Sixty microliters of Diagenode DiaMag protein A/G-coated magnetic beads was washed twice in 200 μL freshly made DiaMag beads wash buffer and then resuspended in 1 mL of DiaMag beads wash buffer and rotated at 4 °C overnight. Blocked protein beads were added to the sample lysates and incubated with rotation at 4 °C for 3 h.
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