![]() Make sure that the methanol concentration in the transfer buffer is not more than 10–20% and that high-quality, analytical grade methanol is used. Thermo Scientific Spectra Multicolor Broad Range Protein Ladder is a 4-color protein standard containing 10 prestained proteins (10 to 260kDa) for use in gel electrophoresis and Western blotting. It may cause a reduction in pore size, precipitation of some proteins, and some basic proteins to become positively charged or neutral. SuperSignal West Femto Maximum Sensitivity Substrate. Methanol removes the SDS from SDS-protein complexes and improves the binding of protein to the membrane, but has some negative effects on the gel itself, leading to a decrease in transfer efficiency. The Color Prestained Protein Standard may also be used in fluorescent imaging of SDS-PAGE.We recommend pre-equilibrating the gel in 2x Transfer buffer (without methanol) containing 0.02–0.04% SDS for 10 minutes before assembling the sandwich and then transferring using 1x transfer buffer containing 10% methanol and 0.01%SDS. For proteins that are difficult to elute from the gel such as large molecular weight proteins, a small amount of SDS may be added to the transfer buffer to improve transfer. I used your Thermo Scientific Unstained Protein MW Marker for a western transfer and got very poor transfer onto the membrane. This inhibition is higher for nitrocellulose than for PVDF. Add freshly prepared DTT solution to a final concentration of 50 mM. SDS in the gel and in the SDS-protein complexes promotes elution of the protein from the gels but inhibits binding of the protein to membranes.Increase voltage, current or length of time for transfer Pre-stained Protein Ladder that can allows for easy identification of clear 12bands between 10 kDa to 245 kDa. Protein ladders and standards for SDS-PAGE, western blots, and isoelectric focusing (IEF) Choose from a variety of protein ladders (molecular weight markers) for protein electrophoresis and western blotting applications.
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