Briefly, 6 mm diameter samples were cut from samples containing different SWNT concentrations, washed with deionized water, sterilized in 70% ethanol for 1 h, and washed thoroughly in sterile Krebs–Henseleit buffer solution (119 mM NaCl, 4.7 mM KCl, 25 mM NaHCO3, 2.5 mM CaCl2, 1.2 mM KH2PO4, 1.2 mM MgSO4) prior to incubation in 10 mL of Krebs–Henseleit buffer solution (pH 7.4).
Samples were hydrated with 100% to 80% ethanol concentration washed with water and the antigen was unmasked by microwaving 20 minutes in the retrieval buffer.
In brief, A549 cells treated with vehicle DMSO or AMEAE at different concentrations were washed with PBS and lysed in ice-cold Radio Immuno Precipitation Assay (RIPA) buffer.
Cells treated with CPP-EGFP or control EGFP at indicated concentrations were washed with HBSS (Invitrogen), with the acid wash buffer (0.2 M glycine, 0.15 M NaCl, pH 3.0) for 30 sec [ 44], and then with HBSS again.
Proteins were precipitated using 5% (final concentration) TCA, washed with acetone, dissolved in 50 mM Tris HCl (pH 8.1) containing 1 mM EDTA and 1% SDS and immunoprecipitated with anti-HA antibody, as described previously (Akiyama et al., 2004).
For survival analysis, cells in mid log phase were incubated for 1 h with H2O2 (at the indicated concentration), washed twice with BG11, plated on solid BG11, and the colonies were counted after 5 7 days of incubation under standard conditions.
In a comparison of hair concentrations between hair washed with only hot water and hair washed once or twice with shampoo, washing hair with shampoo decreased the levels of PCBs, pesticides, and lipids by 25 33% on average (Table 2).
For TUNEL assay, embryos were fixed with 4% paraformaldehyde overnight, rehydrated with decreasing concentrations of methanol, washed with PBST PBSS containing 0.1% Tween 20), treated with proteinase K (10 mg/ml) at 37°C for 30 minutes, refixed with 4% paraformaldehyde, washed in PBST, and lastly incubated with 0.1 M citrate solution (0.1% citrate in PBST).
The concentration bottle was washed with 5 mL sodium dihydrogen phosphate solution and 5 mL MeOH and applied to the SPE column.
Cultures were adjusted so as to contain the same concentration of cells, washed with water, and spotted on plates.
Figure 6b shows the difference between the local protein concentration between resin washed with elution buffer (Protein A only) and washed with binding buffer (Protein A + mAbs, fuchsia marker outline).
