Prestained Protein Ladder is a 3-color protein molecular weight standard containing 10 prestained recombinant proteins covering a wide range molecular weights from 10-170kD. The ladder contains one orange reference band of ~70kD and one green reference band of 10kD., Lot-to-lot variation of the apparent molecular weight of prestained proteins is ~5%. The ladder is supplied in gel loading buffer and is ready-to-use: no heating, further dilution or addition of a reducing agent is required. Prestained Protein Ladder for 100 mini gel applications 5ul per well or 50 large gel applications 10ul per well. 5ul of Prestained Protein Ladder provide 10 bands of equal intensities in 4-20% SDS-PAGE (Tris-glycine buffer) and after electrotransfer onto PVDF membrane. , Applications:,  Monitoring of protein separation during SDS-PAGE (1),  Verifying Western transfer efficiency (2, 3),  Approximate sizing of proteins on SDS-polyacrylamide gels and Western blots,  Locating a protein of interest for excision from an unstained preparative gel. Storage Buffer: 62.5mM Tris-H3PO4 (pH 7.5, 25°C), 1mM EDTA, 2% (w/v) SDS, 10mM DTT, 1mM NaN3, 33% (v/v) glycerol, Recommendations for Loading: 1. Thaw the ladder at RT for a few minutes to dissolve precipitated solids. Do not boil!, 2. Mix gently, but thoroughly, to ensure the solution is homogeneous. , 3. Load the following volumes of the ladder on an SDS-polyacrylamide gel: 5ul per well for mini gel, 10ul per well for large gel. Use the same volumes for Western blotting. The loading volumes listed above are recommended for gels with a thickness of 0.75-1.0mm. The loading volume should be doubled for 1.5mm thick gels. Important Notes: 1. Each lot of the Prestained Protein Ladder is calibrated against Unstained Protein Ladder and calculated apparent molecular weights are reported in the picture., 2. Restained proteins can have different mobilities in various SDS-PAGE-buffer systems. However, they are suitable for approximate molecular weight determination when calibrated against unstained standards in the same system. See reverse page for migration patterns in different electrophoresis conditions., 3. For precise molecular weight determinations use Unstained Protein Ladder. In 8 or 10% gels low molecular weight proteins may migrate with the dye front., 4. Prestained Protein Ladder can be used in Western blotting with all common membranes: PVDF, nylon and nitrocellulose. Longer transfer times or higher transfer voltages may be required, for Western blotting of large (>100kD) proteins. Migration Patterns of Prestained Protein Ladder: This product is manufactured under the license for Strep-tag? technology covered by US patents Nos. 5,506,121, 6,103,493 and foreign counterparts. PRODUCT USE LIMITATION, This product is developed, designed and sold exclusively for research purposes and in vitro use only. The product was not tested for use in diagnostics or for drug development, nor is it suitable for administration to humans or animals. Please refer to www.thermoscientific.com/onebio for Material Safety Data Sheet of the product., © 2012 Thermo Fisher Scientific Inc. All rights reserved. NuPAGE is a registered trademark of Invitrogen. Strep-tag is a registered trademark of Institut fur Bioanalytic GmbH. All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries.