100 mg of purified whole cells is suspended in 2 ml of phosphate
buffered saline and probe sonicated using a Sanyo (MSE) Soniprep 150 Ultrasonic
Homogenizer for 40 one minute cycles. An acid fast stain of
the sonicate is performed to confirm >95% breakage of the cells.
Protein content is quantitated by the BCA assay (Pierce) and
the sonicate is aliquoted at 250 micrograms per vial, luophilized, and stored
at -80oC.
Notes:
This crude
preparation contains proteins, lipids and carbohydrates present
within the bacterial cell, including cell wall, cytosol, and membrane.
