ROTI®Load 1 (reducing) as well as ROTI®Load 2 and 3 (non-reducing) 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.
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.
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.
ROTI®Load 1 is suitable for all PAGE standard gels. It has denaturing and reducing properties.
ROTI®Load 2 is suitable for protein complexes, secondary structures and native gels. It has denaturing properties, but it does not have reducing properties. It is possible to enrich the buffer solution with reducing agents such as β-Mercaptoethanol or DTT.
ROTI®Load 3 is suitable for native gels, precast gels and cooled gels. The buffer solution contains LDS. Unlike SDS, LDS does not crystallise out at low temperatures. In addition, the buffer solution is optimally adapted for precast gels. ROTI®Load 3 has denaturing properties, but does not have reducing properties. It is possible to enrich the buffer solution with reducing agents such as ß-Mercaptoethanol or DTT.
Glycerine is the standard reagent for increasing the density of samples.
SDS (sodium dodecyl sulphate), LDS (lithium dodecyl sulphate) are anionic surfactants. They form a negatively charged mantle around the protein molecules, which results in a denaturing effect. This process is reversible. Irreversible denaturation only occurs if the samples are boiled. Negative charging allows the molecules to be separated during electrophoresis.
β-mercaptoethanol, DTT (dithiothreitol) are reducing agents which split disulphide bridges in protein molecules. Increased unfolding of the molecule occurs. This process is reversible.
Appearance | dark blue, clear solution |
pH value (1x sol.) | 6.6-7.2 |
Remark: Contains SDS. Can be redissolved by heating (max. 45 °C). |