Whole Cell Lysate

Reagent:
Whole Cell Lysate, WCL

Default Quantity:
10 mg

Production system:
Each strain is grown to late-log phase (day 14) in glycerol-alanine-salts (GAS) medium, washed with PBS pH 7.4., and inactivated by gamma-irradiation. Cells are suspended (2 g/ml) in PBS buffer containing 8 mM EDTA, proteinase inhibitors, DNase and RNase. Cells are disrupted by French Press until ~ 90% breakage is obtained (monitored by acid fast staining). The lysate is centrifuged at 3,000 x g to pellet unbroken cells, and the supernatant isolated. The protein content is quantified using the BCA protein assay, and aliquots are stored at -80°C.

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

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