Purified ANX protein is then added in the setting of a sodium borate solution to facilitate covalent interaction of ANX with SPIO in a reducing buffer.
The routine PAGE method utilises a reducing buffer such as SDS and DTT [ 17].
After rinsing with ice-cold PBS, the GST fusion protein was pooled with a reducing buffer of 50 mM Tris-HCl and 10 mM reduced glutathione\ pH 8.0).
The samples were boiled at 100°C for 5 min in a reducing buffer (125 mM Tris-HCl; 140 mM SDS; 20% (v/v) glycerol; 200 mM dithiothreitol (DTT) and 30 mM bromophenol blue) or left at room temperature for 5 min in a non-reducing buffer (as for reducing buffer minus DTT) prior to loading.
Protein samples (30 μg) were denatured in a reducing buffer (62 mmol/L Tris pH 6.8, 10% glycerol, 2% SDS, 5% β-mercaptoethanol, and 0.003% bromophenol blue) and separated by electrophoresis on a sodium dodecyl sulphate (SDS) polyacrylamide gel (10% or 12%).
Subsequently, the pSS patients also had a significantly reduced buffer capacity of UPS and a lower degree of saturation with regard to hydroxyapatite (DSHAP) in their UPS than the controls.
The new process eliminated a unit operation, improved the final concentration of assembly products by a factor of five and reduced buffer consumption nine-fold.
SMB-performance showed reduced buffer consumption, higher purification fold, a better yield and higher productivity.
To further characterize NLRP4 PYD multimerization in solution, we placed the N-labeled NLRP4 PYD in a weakly reducing buffer (0.5 mM TCEP; the NLRP4 PYD has one cysteine, Cys85), waited several days to allow for any dimerization to occur, and recorded a 2D H N HSQC NMR spectrum.
Here, we submerged and boiled the PerioPaper strip in a detergent-free, reducing buffer, added trypsin, and collected liberated peptides after overnight incubation.
