The eluates were pooled and protein refolded by dialysis in refolding buffer with a series of urea buffer for 24 h each (0.05 M Tris pH 8.0, 0.005% triton x-100, 2 mM GSH, urea [6M, 4M, 2M, 1M, 0]).
The capture was performed according to the manufacturer's protocol with streptavidin-coated Dynal beads (Invitrogen, Paisley, UK), and captured samples were washed three times using SureSelect wash buffers with a series of incubation steps.
ATP-injected SNpc tissue blocks (smaller than 1×1×1 mm3) were collected and post-fixed overnight at 4°C using Karnovsky's fixative solution (2% paraformaldehyde, 2% glutaraldehyde, 0.5% calcium chloride in cacodylate buffer, pH 7.2), washed with cacodylate buffer, dehydrated in a series of graded ethanol washes, and embedded in Epon mixture.
After animals were sacrificed, primary tumors were excised and fixed in 4% buffered formaldehyde for 24 h, rinsed with phosphate buffer, dehydrated in a series of graded ethanol and embedded in paraffin.
Agarose beads containing the immune complexes were washed two times each with a series of buffers consisting of the deoxycholate sonication buffer, high salt buffer, LiCl buffer, and TE buffer.
The embryos were fixed in 4 % paraformaldehyde in phosphate buffer (PB), pH 7.4 (PFA), dehydrated with a series of phosphate-buffered saline + 0.1 % Tween 20 (PBSTw)–methanol solutions (3:1 11 3, 1:3) at room temperature (RT) for 10 min each, and finally stored in 100%% methanol overnight at −20 °C.
The lysates were then diluted (2- to 10-fold) with lysis buffer to obtain a series of different sperm concentrations and a standard calibration curve between the fluorescence intensity versus sperm concentrations.
Pieces of apple fruit tissue 5 mm were fixed in either 2% (v/v) formaldehyde with 0.1% (v/v) glutaraldehyde in 0.1 M phosphate buffer, pH 7.2, or 2% (v/v) formaldehyde with 2.5% (v/v) glutaraldehyde in 0.1 M phosphate buffer, under vacuum for 1 h, washed in buffer, dehydrated with an alcohol series, and embedded in LR White resin [ 66].
This preanalytical processing generally involves capturing nucleic acids on microparticles that are then washed with a series of buffers to desorb and dilute out interfering substances.
The American system of government was established to be reflective of the will of population, but with a series of buffers to guard against mob rule.
Excess background and non-specific binding was minimized by washing the filters at 60°C with a series of buffers (2x SSC, 2x SSC with 0.1% SDS, 1x SSC with 0.1% SDS, and 0.5x SSC with 0.1% SDS).
