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TX-114 Soluble Protein Pool
Reagent:
TX-114 Soluble Protein Pool
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. Cells are broken by French press in 4% triton X-114. The materials is then partitioned at 37°C and centrifuged at 27,000 xg, 25°C. The upper aqueous layer is removed and the triton layer is extracted twice more in the same way. The final triton layer is acetone precipitated to remove the triton. The pellet from the acetone precipitation is phenol extracted, then dialyzed into water. The protein content is quantified using the BCA protein assay and aliquots are stored at -80°C.
Notes:
Larger quantities are available for laboratories needing to purify products from this preparation or for laboratories to use for TLR2 signaling.
References:
Radolf, J.D., et. al. Infect. Immun. 56:490-8.
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