Words similar to orthophosphate

Example sentences for: orthophosphate

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Metabolic labeling of cells and immunoprecipitation was performed essentially as described previously [ 21 30 ] . In brief, confluent monolayers of A549 cells were transfected with appropriate dsRNA (at 20 nM final) as described above, and at 8 hr post-transfection, were washed with a phosphate-free buffer (120 mM NaCl, 5 mM KCl, 1.5 mM MgCl 2 , 0.25 mM CaCl 2 , 25 mM NaHCO 3 , 20 mM HEPES [pH 7.4]) and labeled with 32P-orthophosphate (500 μCi/ml) (Amersham) in Dulbecco-modified Eagle MEM lacking both phosphate and pyruvate.
Mutation of Asn122 to Asp by site-directed mutagenesis destroyed the phosphatase activity, confirming the essential role of this residue of PP1 in catalysis [ 19 ] . PfPP1 was inhibited by NaF, inorganic orthophosphate, and pyrophosphate at respective IC 50 values of 2.5 mM, 10 mM, and 90 μM (data not shown).
To monitor total eIF-2α in the cell, a parallel A549 monolayer was treated identically except that the labeled orthophosphate was omitted.
To assess the guanine nucleotide state of selected mycRab24 proteins, cells were transfected with the indicated constructs and incubated with 32P-orthophosphate for 5 h, starting 24 h after transfection.