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SDS-Soluble Cell Wall Proteins
Reagent:
SDS-Soluble Cell Wall Proteins, SCWP
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. Cell walls are isolated as described (see cell wall fraction for details). The cell wall pellet is extracted with 2% SDS in PBS at room temperature for two hours. The SDS extract of the cell wall is removed by centrifugation and SDS is removed by running the extract over a column packed with Extracti gel (Pierce Chemical). The protein content is quantified using the BCA protein assay and aliquots are stored at -80°C.
Notes:
Minimal SDS concentrations are achieved using Extracti-gel and the amount of residual SDS contamination is determined.
References:
Hirschfield, G.R., et al. J. Bacteriol. 172:1005, 1990.
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