Sentence examples for buffer for a further from inspiring English sources

Following this, the fragments were loaded by incubation in 50 μM of the H2O2-sensitive fluorescent dye 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA, Molecular Probes, Leiden, The Netherlands) for 10 min. After washing in fresh buffer for a further 20 min, the fragments were challenged with various compounds as indicated in the figure legends.

Sections were treated with permeabilization solution (Millipore, Watford, UK) for 15 min and blocking buffer for a further 15 min.

The following day, cells were placed in alkaline buffer (0.3 M NaOH, 1 m M EDTA, pH 13) on ice for 40 min, before being electrophoresed at 23 25 V, 300 mA in alkaline buffer for a further 40 min on ice.

Peptides were resolved over 50 min at a flow rate of 300 nL/min with a gradient of 5 40 % buffer B for 40 min followed by a gradient of 40 80 % buffer B for 5 min and held at 80%% buffer B for a further 5 min.

Pellets were further resuspended in 200 µl of lysis buffer, pulsed for a further 5 cycles on the bead-beater and centrifuged as before, and supernatants were combined to the previous ones.

The next day the guts were postfixed for 30 min in 1% osmium tetroxide in 2% glutaraldehyde in 0.1 M phosphate buffer, and for a further 1 hr in 2% osmium tetroxide.

They were rinsed in sodium cacodylate, scraped and pelleted in 2% agar, then post-fixed in 1% osmium tetroxide for 1 hr, en bloc stained in 2% uranyl acetate in maleate buffer pH5.2 for a further hour, rinsed then dehydrated in an ethanol series and infiltrated with resin (Embed812 Electron Microscopy Science) and baked over night at 60°C.

Then, the membrane was washed three more times with TBST buffer, incubated for a further 3 h with a solution of anti-rabbit IgG Fc) goat IgG conjugated with alkaline phosphatase (Promega), washed as before, and visualized with Western blue stabilized substrate (Promega) in accordance with manufacturers' protocol.

Afterward, slides were removed from the lysis buffer and 1.25 ml of dithiothreitol (DTT) was added to lysis buffer and inverted to ensure mixing; then, cells were incubated in lysis buffer with dithiothreitol for a further 30 min at 4 °C, after which lithium diiodosalicyclate (LIS) was added (4 mM final concentration) and incubated for 1.5 h at room temperature.

For the third stage the solution was changed for fresh 0.625% glutaraldehyde (in PBS buffer) and maintained for a further 48 hours.

Following loading cells were washed in Krebs buffer and incubated for a further 20 min to allow de-esterification of the loaded dye.