3 3 cholamidopropyl dimethylammonio 1 propanesulphonate chaps

Analytical-grade chemicals were utilized. BSA (albumin bovine fraction V, pH 7.0), Immobilon-P membrane, Tween 20, Triton X-100 and Coomassie Brilliant Blue (CBB) R-250 were obtained from Serva (Germany). Gelatin, dithiothreitol (DTT), iodoacetamide, 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulphonate (CHAPS) were from Sigma (Prague, Czech Republic); thiourea, urea and IPG buffer (pH 3–10) were purchased from Amersham Biosciences (Uppsala, Sweden). Protein standards were from Bio-Rad (Hercules, CA, USA), chemiluminescent substrate (SuperSignal, Pierce) was obtained from Rockford (USA) and VectaShield mounting medium for fluorescence with DAPI H-1200 from Vector Laboratories, (Burlingame, CA, USA).

Urea, protease inhibitors, ammonium persulfate (APS), bromophenol blue, glycerol, N,N,N′,N′-tetramethylethylene-diamine (TEMED), sodium dodecyl sulfate (SDS), TRIZMA, 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulphonate (CHAPS), dithiothreitol (DTT), iodoacetamide, ethanol, ammonium sulfate, phosphoric acid, Coomassie G-250 were purchased from Sigma-Aldrich (St. Louis, MO, USA). DC Protein assay kit, acrylamide, agarose, ready-made immobilized pH gradient (IPG) strip were purchased from Bio-Rad (Hercules, CA, USA). Ampholine at pH 3.5–10 and 5–8 were obtained from GE Healthcare (Milan, Italy).