Monoclonal Antibodies to Quantitate
نویسندگان
چکیده
content in L2 lung cells and WI-38 fibroblasts cultivated under hypoxic and normoxic conditions. After 96 h of hypoxic cultivation, PyKi activity was significantly increased in both cell types (L2: normoxia [Po2 = 142 torr], 0.11+0.01 [SD]; hypoxia [Po2 = 14 torr], 0.25 ±0.04 U/,ug DNA, P < 0.01). PyKi content increased proportionately in both cell lines (L2: normoxia, 0.44 +0. 13; hypoxia, 0.94±0.13 ,g enzyme protein/,Lg DNA). Specific activity was not significantly different after 96 h (L2: normoxia, 261±11; hypoxia, 261+14 U/mg enzyme protein). These results indicate that regulation of glycolysis during chronic hypoxia occurs at the level of enzyme content. Chronic 02 depletion leads to either an increased rate of biosynthesis or a decreased rate of biodegradation of PyKi, causing augmented glycolytic capacity. Monoclonal antibodies provide a highly specific, convenient approach to characterizing enzymes, as well as quantitating cellular enzyme content.
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