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![]() Do not heat, dilute, add reducing agent before loading.Īpproximately 0.2~0.4 mg/ml of each protein in buffer (20 mM Trisphosphate pH 7.5 at 25☌), 2% SDS, 1 mM 2-Mercaptoethanol, 3.6 M Urea, and 15% (v/v) glycerol).ĥ μl of 10-180kDa Prestained Protein Ladder resolves 10 bands in 4-20% SDS-PAGE (Tris-glycine buffer) and after Western blotting to PVDF membrane. The system includes plasmids which express 10. The ladder is supplied in gel loading buffer and is ready to use. We have created the Penn State Protein Ladder system to produce protein molecular weight markers easily and inexpensively (less than a penny a lane). Unstained Protein Ladders MagicMark XP Western Protein Standard: 20220: 9: all SDS-PAGE gels: stain of choice : IgG binding sites: PageRuler Unstained Protein Ladder: 10200: 14: all SDS-PAGE gels: stain of choice : Strep-tag II sequence : PageRuler Unstained Broad Range Protein Ladder: 5250: 11: all SDS-PAGE gels: stain of choice. What should I do Here are some suggestions: Make sure that the correct amount of standard is loaded per lane. The 10-180kDa Prestained Protein Ladder is designed for monitoring protein separated during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes(PVDF, nylon, or nitrocellulose) and for approximate sizing of proteins. Proteins are covalently coupled with a blue chromophore except for two reference bands (one green and one red band at 25 kDa and 75 kDa respectively) when separated on SDS-PAGE (Tris-glycine buffer). The 10-180kDa Prestained Protein Ladder is a three-color protein standard with 10 pre-stained proteins covering a wide range molecular weights for 10 to 180 kDa. Apply more for thicker (>1.5 mm) or larger gel. For size determination on western blots, depending on the visualization method being used, the protein ladder can be unstained, prestained, or with a Strep -tag sequence.2~3 μl per well for general Western transfering.3 μl or 5 μl per loading for clear visualization during electrophoresis on 15 well or 10 well mini-gel respectively.
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