Urea cell lysis

1mL PBS—on ice, scrape, pellet by spinning 2K rpm for 5mins @ 4C

Re-suspend in 150uL (pending pellet size) urea lysis buffer (ULB in -20)

Sonicate full power for 1min 30 sec (high)

Pipette to check lysate is not viscous

Spin hard (max) 4C for 5 mins

Transfer sup to new tube

Quant if necesary (via Bradford assay--cant use BCA kit with urea lysis buffer) Add LDS (4x)

150uL sample + 50uL LDS = 200uL 1X LDS sample mixture