Example sentences for: pmsf
How can you use “pmsf” in a sentence? Here are some example sentences to help you improve your vocabulary:
For immunoprecipitations, labeled cellular proteins were extracted with lysis buffer (1% of the indicated detergent, 20 mM Hepes, pH 7.4, 150 mM NaCl, 2 mM PMSF, 10 μg/ml leupeptin, 10 μg/ml aprotinin, 2 mM MgCl2, 0.5 mM CaCl2) for 60 minutes at 4°C.
The Triton-insoluble fractions were then scraped into cold Triton X-100 lysis buffer [50 mM Tris/HCl (pH 7.4); 100 mM NaCl; 50 mM NaF; 5 mM EDTA; 40 mM glycerophosphate; 1 mM sodium orthovanadate; 100 μM PMSF; 1 μM leupeptin; 1 μM pepstatin A; 1% (v/v) Triton X-100] and incubated for 20 min on ice, and clarified by centrifugation at 13000 g for 5 min at 4°C.
Following the 48-72 hour incubation, the media was collected and placed on ice with the following protease inhibitors added: 1 mM DTT (Sigma, St. Louis, MO), 0.5 mM PMSF (Sigma, St. Louis, MO), 5 μg/ml Pepstatin (Sigma, St. Louis, MO), 3 μg/ml Leupeptin (Sigma, St. Louis, MO) and 5 μg/ml Aprotinin (Sigma, St. Louis, MO).
In order to observe changes at the onset of apoptosis, only adherent cell populations were trypsinized, pelleted 5 minutes at 500 × g, and lysed in ice cold lysis buffer (10 mM Tris-HCl (pH 7.5), 1 mM MgCl 2 , 1 mM EGTA, 0.1 mM PMSF, 5 mM β-mercaptoethanol, 0.5% CHAPS, 10% glycerol) for 30 minutes at 4°C.
Next, the cells were resuspended at 3 × 10 6cells/ml in ice cold sonication buffer (250 mM sucrose; 10 mM HEPES, pH 7.4; 1 mM EDTA; 5 mM MgCl 2 ; 1 mM DTT; 0.1 mM GTP; 1 μg/ml leupeptin, 1 μg/ml pepstatin A, and 1 μg/ml aprotinin; 1 mM PMSF) and sonicated for 10 seconds.
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