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Example sentences for: hcl

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The inhibitory activities of fibrosarcoma PAI-1, wtPAI-1, and its mutants were measured by a direct chromogenic assay using uPA (100 U/ug) and its substrate S-2444 in a microtiter plate as described earlier [ 20 ] . Briefly, fibrosarcoma PAI-1, wtPAI-1, or its mutants were diluted 1:10 in the activity measuring assay buffer (50 uM Tris-HCl, pH 7.5, containing 0.15 M NaCl and 100 ug/ml BSA).
Plasma samples 1 μl were added to 2 ml vanadium III chloride (Sigma-Aldrich) dissolved in 1 N HCl at 90°C to reduce all NO metabolites to NO.
Smooth muscle strips were weighed and total protein was extracted by glass-glass homogenization using 50 μl per milligram dry tissue sodium dodecyl sulfate (SDS)-based extraction buffer: 25 mM Tris-HCl, pH 7.4, 2% SDS, 10% glycerol, 1 mM dithiothreitol (DTT), 1 μM leupeptin, 10 mM ethylenediaminetetraacetic acid (EDTA), 1 mM sodium orthovanadate, 5 mM NaF and 1 mM phenylmethylsulfonyl fluoride (PMSF).
Fractions (2 ml) were collected and neutralized with 2 M Tris-HCl.
Duplicates of membrane preparations equivalent to 0.1 mg protein were incubated with 125I-EGF (50 or 100 ng; Amersham Life Sciences, Arlington Heights, IL) in 200 μl reaction buffer (5 mM Tris-HCl, pH 7.0, 125 mM sucrose, 75 mM NaCl, 0.5 mM Ca 2+and 0.5% BSA) at 25°C for 60 min.