Thats mostly what I wanted
10.1016/j.chom.2018.02.004 22 CokerO
The slight warmth and gentle motion will usually coax the remaining lyophilized powder into solution
2.4 Western blot analyses Whole cell protein lysates were prepared by lysis in radio-immunoprecipitation assay buffer (150 mM NaCl, 2 % NP-40, 0.5 % sodium deoxycholate, 0.1 % SDS, 50 mM Tris pH 8.0, 1 mM PMSF, 1 g/ml aproptinin, 1 g/ml pepstatin, 1 g/ml leupeptin, 1 mM NaF, 1 mM sodium orthovanadate, 0.1 mM sodium molybdate, 10 mM sodium pyrophosphate) at a density of 4 10 7 cells/ml on ice, followed by addition of equal volumes of Laemmli protein sample buffer (60 mM TrisCl pH 6.8, 2 % SDS, 10 % glycerol, 5 % -mercaptoethanol, 0.01 % bromophenol blue) and heated to 95 C for 5 min prior to separation on SDS-PAGE gels and transfer to 0.45 m nitrocellulose membranes