Substrate solutions of 1% (w/v) were prepared in a buffer of optimum pH activity for the enzyme.
Samples were prepared in a buffer of 50 mM Tris HCl, 150 mM NaCl, 20 μM EDTA, pH 8.0 and equilibrated at room temperature for at least 30 min before measurements were taken on a two-channel fluorometer (Photon Technology International Alphascan, Birmingham, NJ).
Prior to Western blotting, the sample is prepared in a buffer consisting of 2% SDS and 100 mM of the reducing agent dithiothreitol (DTT) then heated for 3 min at 90 °C.
The solutions of different concentrations of Bi III) ions ranging from 0.001 to 1 ppm were prepared in a buffer solution of pH 4.
Whole-cellular lysates were prepared in a buffer, gentle shaking, composed of 0.9% NaCl, 20 mℳ Tris-ClH, pH 7.6, 0.1% triton X-100, 1 mℳ phenylmethylsulfonylfluoride and 0.01% leupeptine.
Whole cellular lysates were prepared in a buffer, with gentle shaking, composed of 0.9% NaCl, 20 mM Tris-HCl, pH 7.6, 0.1% Triton X-100, 1 mM phenylmethylsulfonylfluoride and 0.01% leupeptine.
DAOY cell lysates were prepared in a lysis buffer of 100 mM HEPES, pH 7.6, 300 mM NaCl, 0.1% Triton X-100, 2 mM EDTA, 2 mM EGTA supplemented with phosphatase and protease inhibitors.
The SWW was prepared in a phosphate buffer (NaH2PO4 of 6 g L−1 and Na2HPO4 of 7.1 g L−1) of pH 7.03 that contains the following nutrients or salts: peptone, 0.12 g; yeast extract, 0.12 g; NaCl, 0.007 g; MgSO4·7H2O, 0.002 g; CaCl2·2H2O, 0.004 g, and 200 µL of a trace metal solution per liter of distilled/deionized water (Bajaj et al. 2008).
The immunological reaction was carried out as follows: Firstly, the six kinds of biomarkers related to gastric cancer (CEA, CA19-9, H.P53 PG3, PG I, and PG II) solutions were respectively prepared in a series of concentration with phosphate buffer solution (PBS, pH = 7.4) (Table. 1).
Briefly, a working solution of 2% annexin V and 1 μg/mL propidium iodide (PI) was prepared in a solution of 1× annexin binding buffer.
The samples were then passed through a 0.2 μm PTFE syringe filter and diluted in duplicate 10-, 100-, and 10 000-fold into a mixture of 1 1 buffer: acetonitrile prior to analysis, and assayed by LC-MS/MS using electrospray ionization against standards prepared in a mixture of 1 1 assay buffer/acetonitrile.
