The supernatant was taken out and mixed with the same volume of loading buffer and then was analyzed with SDS-PAGE.
AS/RS structures are designed according to their different size, weight, and volume of loads to be handled, and characterized of the warehouse.
A 0.1* volume of loading buffer (containing 0.3% bromophenol blue, 50% glycerol, 0.3% mercaptoethanol, and 50% [v/v] lysis buffer) was added.
For HIC, two 5-mL HiTrap Octyl FF columns (GE Healthcare) were serially connected and equilibrated with one column volume of loading buffer (10 mL of 200 mM ammonium sulphate in 50 mM Tris-HCl buffer, pH 7.0).
Pellet volume was estimated and re-suspended into twice the volume of Loading Buffer.
The reactions were stopped by the addition of one volume of loading buffer, fractionated on denaturing 5 8% PAGE gels which were then exposed to PhosphorImager screens.
Incubation was terminated by addition of an equal volume of loading dye-urea-TTE and electrophoresis on a 20% denaturing polyacrylamide gel.
An equal volume of loading dye-urea-TTE was added and products were fractionated by electrophoresis through a 20% denaturing polyacrylamide gel.
Protein lysate was denatured in an equal volume of loading buffer (100 mM Tris-HCl, pH 6.8, 200 mM DTT, 4% SDS, 0.2% bromophenol blue, 20% glycerol), heated at 95°C for 5 minutes, and separated on 10% SDS-PAGE.
For Western blot analysis, the soluble and insoluble protein preparations were mixed separately in the same volume of loading buffer, separated on SDS/PAGE gels and then transferred onto nitrocellulose membrane.
Samples were diluted with one volume of loading dye (95% formamide, 0.05% xylene cyanol FF, and 0.05% bromophenol blue), heat denatured at 95°C for 5 min, and immediately cooled on ice.
