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Additional data should be considered for a more accurate adjustment.Ġ.2~0.4 mg/ml of each band of protein is stored in a mixture of solution that contains: 20 mM Trisphosphate at pH 7.5, 2% SDS, 0.2 mM Dithiothreitol, 3.6 M Urea, and 15% (v/v) Glycerol.ĥμl of IRIS9 Prestained Protein Ladder (PM009) resolves 9 bands in 4-20% SDS-PAGE (Tris-glycine buffer) and after Western blotting to PVDF membrane. Note: The molecular weight of each protein (kDa) was measured against an unstained protein ladder in every electrophoresis condition. Use Precision Plus Protein WesternC Standards as a combination standard for western blotting and fluorescence detection in SDS-PAGE. The protein ladder is supplied in a ready-to-use format for direct loading onto gels no need to heat, reduce. 2.5 μl per well for general Western transferring. Thermo Scientific PageRuler Unstained Protein Ladder is a mixture of 14 recombinant, highly purified, unstained proteins (10 to 200 kDa) for use as size standards in protein electrophoresis (SDS-PAGE) and western blotting. Refer to the IRIS9 Prestained Protein Ladder patterns in various electrophoresis conditions:ģ μl or 5 μl per loading for clear visualization during electrophoresis on 15-well or 10-well mini-gel, respectively. Guide for Molecular Weight Estimation (kDa) The smaller the size of the protein of interest, the higher the percentage of acrylamide/bis. The example shows a visible image (left) and a two-color, near-infrared image (right) resolved on a 4 - 12 Bis-Tris Gel and transferred to nitrocellulose via wet tank transfer. The ScanLater Western Blot Protein Ladder consists of seven biotinylated recombinant proteins at 10, 20, 40, 50, 80, 100 and 140 kD. one plane of separation) or two dimensional. In blotting applications, the ladder can be used to monitor protein transfer and as a reference to estimate the molecular weight of proteins of interest. Electrophoresis can be one dimensional (i.e. The IRIS9 Prestained Protein Ladder keeps track of the size and separation of proteins during SDS-polyacrylamide gel electrophoresis, approximating the target protein size and validating the Western transfer efficiency on PVDF, nylon, or nitrocellulose membranes. Download the complete western blot guide. The 9 recombinant proteins are covalently coupled with blue chromophore, while 2 red bands at 70 kDa and 30 kDa, a green band at 15 kDa and a newly designed peacock green band at 60 kDa serve as reference bands. The IRIS9 Prestained Protein Ladder is a combination of 9 pre-stained proteins with molecular weights from 15 to 180 kDa. Monitoring protein transfer onto membranes after western blotting Sizing of proteins on SDS-PAGE gels and western blots.