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Example sentences for: s-
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After transient expression of the α subunits, cellular proteins were radiolabeled with a brief pulse of 35S-methionine followed by a chase.
This does not lead to a major drop in total protein synthesis as indicated by metabolic labeling with 35S-amino acids.
Proteins resolved by SDS-PAGE were transferred to Immobilon-P membranes (Millipore Corp., Bedford, MA) using carbonate transfer buffer (pH 9.9) [ 5 ] . Specific proteins were visualized using rabbit polyclonal antibodies specific for glutathione S-transferase (GST)-tagged LcrG (α-LcrG) [ 4 ] , His-tagged LcrV (α-LcrV) [ 4 ] , YopM (α-YopM) [ 36 ] , GST-tagged LcrQ (α-LcrQ) [ 16 ] , YopN (aka LcrE) (α-LcrE [ 4 ] ), His-tagged YopD (α-YopD) [ 16 ] , and YopE (α-YopE; gift from S. C. Straley, University of Kentucky, Lexington, KY.
Primary fibroblast cultures from patients and control individuals were grown to confluency and then metabolically labeled with 35S-cysteine as described [ 17 18 ] . The soluble cell lysis fraction, containing newly synthesized labeled fibrillin-1 molecules, and the insoluble fraction, representing the labeled fibrillin in the extracellular matrix, were fractionated by SDS polyacrylamide gel electrophoresis and autoradiographed.
Sodium salts of each compound were titrated into assays of purified Fhit [ 16 ] with 1.8 μM fluorescent substrate, Appp-S-(4-4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacine-3-yl)methylaminoacetyl (ApppBODIPY) [ 10 ] , and competitive K i values were obtained by calculating the inhibitor concentration-dependence in reduction of k cat / K m (apparent) for substrate hydrolysis [ 10 ] . As shown in Table 1, inhibitors 2b and 3b with two-carbon diol linkers were either only as inhibitory as the natural substrate 1 or substantially less so.
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