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
preparation contains proteins, lipids and carbohydrates present
within the bacterial cell, including cell wall, cytosol, and membrane.