The resulting slurry was transferred to a gravity-flow column and washed twice with 50 ml of lysis buffer.
The beads were washed four times with 1 ml of lysis buffer and then subjected to SDS polyacrylamide gel electrophoresis analysis.
Cells were harvested by centrifugation (5,000 × g for 5 minutes) and resuspended in a small amount (typically 0.2 mL) of lysis buffer (50 mM Tris, pH 8.0, 0.1 mM benzamidine, 0.5 mM EDTA).
The cells were harvested, resuspended in 50 ml of lysis buffer (50 mM PB [pH8.0], 200 mM NaCl, 1 mM DTT, and one tablet of Complete TabletTM (Roche)) per 1 l culture, and then disrupted using Bioruptor (COSMO BIO).
Bacterial pellets were lysed in 30 mL of lysis buffer (100 mM KCl, 20 mM HEPES-KOH, pH 7.5), and homogenized by sonication (Sonifier S-250A analog ultrasonic processor).
Thereafter, the beads-antibody-antigen complex was pelleted and washed 3 times with 1 mL of lysis buffer.
Beads were washed twice with 1 ml of lysis buffer.
The immunoprecipitates were washed 3X with 1 ml of lysis buffer.
We washed the beads five times with 1 mL of lysis buffer.
The sepharose beads were washed three times with 1 ml of lysis buffer.
The cells were harvested by centrifugation and resuspended in 30 ml of lysis buffer.
