THP-1 cells were seeded at 0.2x[10.sup.6] cells/ml in 25 ml of routine culture medium containing HS and 2-mercaptoethanol
(as described in Section 2.2) and cultured for either 48 or 72 [+ or -]2 h prior to dosing with proficiency substances.
EDC (2.0 mg) and sulfo-NHS (5.5 mg) were added to PEG-USPIO in 0.1 M MES buffer solution, and the reaction was maintained for 15 minutes at room temperature (RT) and then quenched with 2-mercaptoethanol
. The solvent was removed by centrifugation at RT for 20 minutes at x2500g (Millipore Amicon Ultra, Massachusetts, USA), and the resultant NPs were resuspended in PBS buffer solution and mixed with SA (3.0 mg SA) for 2 hours while stirring at RT before the reaction was stopped by adding ethanolamine.
Weight percent Nanocomposite acrylic resin Raw materials Acrylic resin a b Methyl methacrylate 20 19.8 19.5 Butyl acrylate 22.5 22 21.5 Styrene 13 12.7 12.5 Acrylic acid 1 1 1 2-Mercaptoethanol
0.5 0.5 0.5 Dibenzoyl peroxide 3 3 3 Cloisite 30B -- 1 2 Solvent 40 40 40 Table 3: Traffic marking paint formulations.
Effect of salts and other effectors on the protease activity: The effect of different salts (MgS[O.sub.4], ZnS[O.sub.4], CuS[O.sub.4], NaCl, and KCl) and other effectors (EDTA, 2-mercaptoethanol
, sodium thiosulfate) at different concentration was measured.
Determination of IgG Levels: Same protocol was adopted for IgG titers but in the first step 50 u l of 0.01M 2-mercaptoethanol
(Riedel-de Haen, Germany) was added in phosphate buffered saline in each well that destroyed the IgM immunoglobulins and the remaining titer was of IgG.
Briefly, cells from bone marrow flushed from femurs and tibiae of mice were cultured in 6-well tissue culture plates at 1 x [10.sup.6] cells/ml in complete RPMI culture medium supplemented with 10% FBS and 2-mercaptoethanol
(50 [micro]M/ml) in the presence of IL-4 (1000 U/ml) and GM-CSF (1000 U/ml).
, sodium pyruvate, glucose, MDC (monodansylcadaverine), and mouse anti[beta]-actin monoclonal antibody were obtained from SigmaAldrich (St.
Sodium pyruvate, HEPES, 2-mercaptoethanol
, dimethyl sulfoxide (DMSO), 3-isobutyl-1-methylxanthine (IBMX), Forskolin, thiazolyl blue tetrazolium blue, Hoechst 33342, and propidium iodide were purchased from Sigma-Aldrich (MO, USA).
Synthetic glycopeptides and glycopeptides obtained from double proteolytic digestion were mixed with 1 [micro]L 2-mercaptoethanol
and dimethylamine to give a final concentration of 30% and incubated at 55 [degrees]C for 6 h.
The inhibitor used were EDTA, 2-mercaptoethanol
and inducer was copper sulphate.
, acetic acid, hydrochloric acid, phosphoric acid, casein, gelatin, tyrosine, 2-propanol, leupeptin, phenylmethylsulfonyl fluoride (PMSF), sodium dodecyl sulfate (SDS) and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St.
Shoots and roots frozen in liquid nitrogen were ground into a powder and then extracted with 0.2 mol*L-1 Tris-HCl buffer (pH 7.2) containing 5 mmol*L-1 2-mercaptoethanol
, 1 mmol*L-1 ethylene diamine tetraacetic acid, and 7.5% polyvinylpolypyrrolidone.