Preparation of E. coli lysate with B-PER solution


1 Induce expression of E. coli strain BL21 (DE3) or other suitable expression host containing gene of interest
2 Centrifuge at 10,000 x g for 10 minutes at 4°C
3 Discard supernatant and freeze pellet at -70°C
4 Add 5 ml B-PER (Thermo) per gram of pellet and resuspend pellet
5 Incubate with rotation for 15 minutes at Room Temperature
6 Centrifuge at 30,000 x g for 10 minutes at 4°C
7 Transfer supernatant to 50 ml tube and adjust lysate to less than 20% BPER with MBS or PBS
Lysate is now ready for incubation with nanoCLAMP(Resin)

Solution(s) Used
Use / Nickname Solution
MBS MBS pH 6.5

Outside Material(s) Used
Vendor Outside Material Catalog Number Usage Rate
ThermoFisher Scientific B-PER 78248 5 ml per per g pellet