To answer this question we have carried out thermal denaturation studies of lysozyme, ribonuclease-A, and apo-alpha-lactalbumin in the presence of various TMAO concentrations at different pH values above and below the pK(a) of TMAO. The main conclusion of this study is that near room ...
pKa
A 1 ml aliquot was stored at -20 °C, and bacteria were pelleted at 8,000xg for 20 min at 4 °C and resuspended in 8 ml native buffer (50mM N aH2PO4, 300mM NaCl, 20mM imidazole, pH 8); lysozyme was added to 1 mg/ml and the resuspension was incubated on ...
Lysozyme Solution 10 mg of lysozyme 10 ml of H2O Glutathione Elution Buffer 40 mM reduced glutathione 200 mM NaCl 0.2% Tween 20 100 mM Tris-HCl, pH 8.5 Sodium dodecylsulfate (SDS) Sample Buffer 100 mM Tris-HCl, pH 6.8 4% SDS 20% glycerol 10% β-Mercaptoethanol 0.02% bromophenol blue ...
Cultures were grown for 20-24hrs at 20°C. Cultures were harvested by centrifugation at 4000x g, removal of media, and frozen at −80°C for future analysis. Protein Purification Cells were lysed in buffer containing 10% glycerol, 1 mg/mL lysozyme, 50mM Tris-HCL pH 7.8, 500mM NaCl,...
Cells were lysed via sonication in 10 mM HEPES pH 7.5 and 250 mM KCl (buffer A), 5 mM MgCl2, and 10 mM Imidazole, supplemented with 25 mg/ml DNaseI, 25 mg/ml lysozyme, and fresh phenylmethylsulfonyl fluoride (PMSF) to a final concentration of 0.1 mM. Phosphorylation loop of RyR dom...