ROTI ® Load 1 (reducing) as well as ROTI ® Load 2 and 3 (non-redu cing) represent special gel loading buffers for protein gel electrophoresis. They have been designed to protect your proteins during sample preparation and to stabilise sensitive peptide bonds. Protein degradation is efficiently prevented by those buffer systems. ROTI ® Load are 4x concentrated ready-to-use solutions, which may be used directly for sample loading. ROTI ® Load 1 4x conc., reducing Well tested and proven sample buffer for protein gel electrophoresis that has been modified and further developed based on the original formulation acc. to Laemmli ( Nature 227, 1970). Optimized for a distinct band display. Recommended for size estimation of proteins in gel. Formulation acc. to Laemmli (modified) Stabilises peptide bands Prevents protein degradation Improves gel separation Directions for use ROTI ® Load protein gel loading buffers are usually used as 1x concentrated solution. ROTI ® Load can either be added to the prepared sample or the precipitated proteins are dissolved directly into ROTI ® Load. Heat the ROTI ® Load mixed sample to 95 °C for 5 minutes to break hydrogen bonds and stretch the peptide chains. The β-Mercaptoethanol contained in ROTI ® Load 1 opens disulfide bridges by reduction and leads to a complete molecular stretching, the basic requirement for a correct size estimation of proteins in the gel. ROTI ® Load 1 prevents degradation of proteins while heating during sample preparation, additionally keeping the sample pH value constant during gel electrophoresis. Being a phosphate-buffered ready-to-use solution, ROTI ® Load 1 already contains all necessary denaturing and reducing reagents, bromophenol blue as dye and glycerol as density increasing reagent.