DSP crosslinking (on plate)
Plated cells (on 10cm diameter dishes)
Discard medium
2 washes in PBS
Add 2ml of 1mM DSP in PBS, 250mM sucrose (starting from a 100X DSP solution in DMSO)
30’ at 4°C on rotating platform
Discard DSP
1 wash in PBS 20mM TrisHcl pH 7.4 (quencing)
2 washes in 2 ml PBS 20mM TrisHcl pH 7.4 (quencing), 2ml/plate, 15’ at 4°C on rotating platform
Then lysis: 1 washes in PBS 10mM NEM
Lysis in RIPA buffer on the plate (1ml buffer/plate) with scaper