After electrophoresis, gels were immediately washed with distilled water and activity staining of bacterial PO was conducted in 100 mM NAc buffer (pH 5) containing different PO substrates such as 2,6-DMP, ABTS, guaiacol and para-phenylenediamine.
Plant part samples (roots, twigs, bark, phloem, wood, leaves) collected from mature plants of the new Actephila were immediately washed with demineralised water, oven-dried at 70 °C and then digested with concentrated nitric acid (70 %) and hydrogen peroxide (30%%) in a specialised microwave.
The petioles were immediately washed with water, then wrapped with cotton cord.
Cells were immediately washed with ice cold phosphate-buffered saline (pH 7.4).
For antigen retrieval, OB slices were steamed for 10 minutes in a solution of 0.01 M Sodium Citrate, then immediately washed with PBS-T.
After being treated with Berberine or vehicle, HUVEC 100-cm22 dishes) were immediately washed with ice-cold PBS, then scraped with HClO4, and centrifuged at 4°C (5 min, 14,000 rpm).
2.5 3.0 mm with uniform shape were selected and immediately washed with plenty of deionized water until the solution became neutral, and then stored in water at 4°C till activation.
For total protein extraction, tissues were immediately washed with PBS after dissection and lysed in S1 buffer (50 mMTris, pH 8; 0.4% NP-40; 150 mMNaCl; 1 mM EDTA; proteinase inhibitor cocktail tablet (Roche); 1 mM PMSF; 2 mM N-ethylmaleimide), using a pestle for homogenization.
At that time cells were washed with 2x SSC, treated with 0.5% saponin/0.5% Triton X-100 in 2x SSC, 1 hr, followed by 1M HCl for 5 min. They were then immediately washed with 2xSSC three times before the buffer was replaced to TN-Buffer (0.1 M Tris-HCl pH 7.5+150 mM NaCl) for 10 min, then blocked by TNB (TN-Buffer with 0.5% Blocking reagent) (Perkin Elmer, FP1020) at room temperature.
The cavity was immediately washed with sterile PBS.
