Two plates of wildtype H. pylori or arsR(D52N) were suspended in 7 mL BHI and 3 mL of each suspension was added to a dialysis cassette as described above. Bacteria were incubated in the dialysis cassettes suspended in 1500 mL of media at pH 3.0 or 7.4 with 5 mM urea for 4 hours. Bacteria were then pelleted at 10,000 rcf, 4°C for 5 minutes. Pellets were resuspended in 3 mL ice cold 25 mM phosphate buffer with 5 mM EDTA and 150 μL bacterial protease inhibitor (Sigma). Bacteria were lysed by 3 passes through a French Press at 20,000 p.s.i. Unbroken bacteria were harvested by a 10 minute spin at 4°C, 1500 rcf. The supernate fraction was collected as total bacterial lysate. Protein concentration was determined by the BCA method (Pierce/Thermo).
Bacterial protease inhibitor
Manufactured by Merck Group
Sourced in Australia
Bacterial protease inhibitor is a lab equipment product that functions to inhibit the activity of proteases derived from bacterial sources. It is used to prevent the degradation of proteins in biological samples by bacterial proteases.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bacterial protease inhibitor
1
H. pylori Protein Extraction Protocol
2
Vaginal Fluid Collection for Analyses
Check if the same lab product or an alternative is used in the 5 most similar protocols
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LASIK PVA eye sponges (Visitec, USA) were placed in the posterior fornix of the vagina for 2 min to absorb secretions. Sponges were immediately placed at −20°C until vaginal fluids were eluted at the same day, as previously described by Kozlowski et al. (2000 (link)), using 300 μl of PBS containing 1% Bacterial Protease Inhibitor (Sigma, Australia) and 10% igepal (Sigma, Australia).
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