It is concluded that the estimated molecular weight of SMOBIO’s pre-stained marker shows a curve matching well with that of unstained native proteins (MARK12), representing a good estimation of the MW of each pre-stained protein in the SDS-PAGE analysis. Although it is impossible to define "precision" for molecular weight of proteins in SDS-PAGE, we did compare the migration pattern of pre-stained markers with unstained protein marker (Invitrogen MARK12) for calibration. Therefore, in the product description, we suggest our users to calibrate the MW against their interested proteins. We did compare the migration patterns of SMOBIO’s Protein Markers with other brands, and we concluded that it was difficult to define “precision” due to the reasons mentioned above. For example, a protein which is highly hydrophilic might show a particular higher position in the SDS-PAGE analysis when compared to a hydrophobic one. It is known that the analysis of protein size by an SDS-PAGE is only for “estimation” because of the intrinsic variation of amino acid composition in all proteins including stained and non-stained ones. Usually, pre-stained marker is written on “estimated molecular weight” for caution. MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane. Under suggested conditions, PM5200 ExcelBand™ 3-color Pre-Stained Protein Ladder Broad Range resolves 15 major bands in SDS-PAGE ( Tris-Glycine, Three reference bands - 75 kDa (red), 40 kDa (green), and 20 kDa (blue)Īpproximately 0.1~0.4 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25☌), 2% SDS, 0.2 mM DTT, 3.6 M urea, and 15% (v/v) glycerol). Ready-to-use - Premixed with a loading buffer for direct loading, no need to boil. The PM5200 3-color Pre-Stained Protein Ladder Broad Range is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins. Proteins are covalently coupled with different chromophores for easy identification of bands, with three reference proteins carrying enhanced intensity corresponding to a blue band at 20 kDa, green at 40 kDa, and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). D-2010, D-2020, and D-2030) are compatible with those staining.The PM5200 3-color Pre-Stained Protein Ladder Broad Range is a ready-to-use three-color protein standard with 15 pre-stained proteins covering a wide range of molecular weights from 5 to 245 kDa in Tris-Glycine Buffer (3.5 to 235 kDa in Bis-Tris (MOPS) buffer and Bis-Tris (MES) buffer).
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