DSP crosslinking

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