Words similar to orthophosphate
Example sentences for: orthophosphate
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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.
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.
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.