Thaw the ladder either at room temperature or at 37-40☌ for a few minutes to dissolve precipitated solids. In this technique a mixture of proteins is separated based on molecular weight, and thus by type, through gel electrophoresis. The quality of the BlueRAY Prestained Protein Ladder is tested on a lot-to-lot basis to ensure consistent product quality.īlueRAY Prestained Protein Ladder Protocolġ. Western blot is often used in research to separate and identify proteins. Make sure sample preparation conditions have not destroyed the antigenicity of the sample. Sizing of proteins on SDS-PAGE gels and Western blots.Īpproximately 0.2~0.4 mg/ml of each protein in buffer (20 mM Trisphosphate pH 7.5 at 25☌), 2% SDS, 0.2 mM Dithiothreitol, 3.6 M Urea, and 15% (v/v) Glycerol. Use molecular weight markers compatible with a western- imaging substrate, such as the Invitrogen iBright Prestained Protein Ladder or Invitrogen MagicMark XP Western Protein Standard, as a positive control. Monitoring of protein transfer onto membranes during Western blots. Monitoring of protein migration during SDS-PAGE. Ready-to-use: supplied in a loading buffer for direct loading on gelsĮasy to identify: includes the ~25, ~75 kDa reference bands coupled with a green and a red dye Do not heats, dilute, and add reducing agent before loading.īroad range: 10-180 kDa (Tris-glycine-SDS running buffer) The ladder is supplied in gel loading buffer and is ready to use. The BlueRAY Prestained Protein Ladder is designed for monitoring protein separated during SDS-PAGE, verification of Western transfer efficiency on membranes (PVDF, nylon or nitrocellulose) and for approximate sizing of proteins. Burnette in 1981 after the eponymous Southern blot for DNA and the consequent coinage of the northern blot in 1977 for RNA.12 Western blotting separates, detects, and identifies one or more proteins in a complex mixture. Protein ladders for IEF, native PAGE and specialty applications. Protein ladders for western blotting and fluorescence western blotting. 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-polyacrylamide gel electrophoresis (SDS-PAGE) with Tris-glycine-SDS running buffer. The name ‘western’ blot was first coined by Dr. On this page: Prestained broad and high molecular weight protein ladders. The BlueRAY Prestained Protein Ladder is a three-color protein standard with 10 pre-stained proteins covering a wide range molecular weights for 10 to 180 kilodalton (kDa).