EVs were resuspended in the lysis solution containing 12 mM sodium deoxycholate (SDC; Sigma-Aldrich, cat. no. D6750), 12 mM sodium lauroyl sarcosinate (sarkosyl; Sigma-Aldrich, cat. no. L9150), 10 mM TCEP-HCl (Sigma-Aldrich, cat. no. C4706), 40 mM ...
EVs were resuspended in the lysis solution containing 12 mM sodium deoxycholate (SDC; Sigma-Aldrich, cat. no. D6750), 12 mM sodium lauroyl sarcosinate (sarkosyl; Sigma-Aldrich, cat. no. L9150), 10 mM TCEP-HCl (Sigma-Aldrich, cat. no. C4706), 40 mM ...
Synovial mesenchymal stem cells (MSCs) have high freeze–thaw tolerance, whereas human umbilical vein endothelial cells (HUVECs) have low freezing tolerance. The differences in cell type-specific freeze–thaw tolerance and the mechanisms involved are unc
Incubations were performed in 96-well plates with 10 nM antithrombin, 2.0 nM FXa or thrombin (all from Hematologic Technologies; Essex Junction, USA) and 0 to 2 μg mL−1 heparins (dry weight) in TS/PEG buffer (0.02 M Tris/HCl, 0.15 M NaCl and 1.0 mg mL...
Whole cell extracts were prepared using RIPA lysis buffer (10 mM Tris HCl, pH 8.0, 1mM EDTA, 150 mM NaCl, 1% Triton-X-100, 0.1% sodium deoxycholate, 0.1% SDS) in the presence of 1X Halt protease inhibitor (Thermo Fisher) and 1 mM PMSF (Sigma, St Louis, MA). The resulting protein...
Cells were incubated in buffer A (10 mM Tris, 1 mM KCl, 1.5 mM MgCl2) for 10 min on ice then dounce homogenized with 10 strokes. The mixture was centrifuged, and the pellet was resuspended in buffer B (50 mM Tris-HCl, 10 mM EDTA, 1% SDS) and kept for 10 min on ice. DNA was...
The recombinant protein was expressed in SoluBL21 (Genlantis) competent cells hosting pRARE plasmids by following the manufacturer's protocol, then extracted with 20 mM Tris–HCl lysis buffer (pH 7.5), purified by nickel affinity chromatography (Ni Sepharose 6 Fast Flow, GE Healthcare), and ...
For analyses of DDR1 expression in whole cell lysates, cultured prostate epithelial cells were lysed in RIPA buffer (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 1% NP-40, 0.25% sodium deoxycholate and 1 mM EDTA) supplemented with protease inhibitors Cocktail, EDTA-free (Cat. # 539134, Sigma-...
(E3889; Sigma-Aldrich), 0.5% Triton-X-100 (Sigma-Aldrich) and 1 mM dithiothreitol (DTT; D0632; Sigma-Aldrich) supplemented with protease inhibitor cocktail (P9340; Sigma-Aldrich) or lysed in 2× sample buffer (125 mM Tris-HCl, pH 6.8, 4% SDS, 20% glycerol, 200 mM DTT and...
U2OS cells carrying latent HHV-6A treated with solvent control or TSA were lysed in RIPA buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 1% Triton X-100, 1% NP-40, 0.1% SDS, 10% Glycerol and protease inhibitor cocktail (Roche)). Protein lysates were incubated with control...