Cell Wall

Reagent:
Cell Wall Fraction, CW

Default Quantity:
1 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. The bacilli are suspended (2 g/ml) in PBS containing 8 mM EDTA, DNase, RNase and a proteinase inhibitor tablet, and broken in a French Press pressure cell at 4°C. Unbroken cells are removed by low speed (3,000 x g) centrifugation. The cell wall is isolated by centrifugation at 27,000 x g for one hour and washed 2 times in PBS. The final cell wall pellet is suspended and dialyzed in 0.01M ammonium bicarbonate, quantified by BCA protein assay for protein content, and stored at -80°C.

Notes:
This preparation contains proteins and non-protein compounds such as mAGP.

References:
Hirschfield, G.R. et al. J. Bacteriol. 172:1005, 1990.

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