![]() This induces the proteins to migrate from the. The gel is placed next to the membrane, and an electrical current is applied. The immunoassay uses a membrane made of nitrocellulose or PVDF (polyvinylidene fluoride). Thaw the ladder either at room temperature or at 37-40☌ for a few minutes to dissolve precipitated solids. Western blotting is a technique that uses specific antibodies to identify proteins separated by size through gel electrophoresis. The detection reagents can be removed and the entire blot reprobed to visualize another protein or to optimize. Chemiluminescent or Fluorescent Western Blotting. Using chemiluminescence allows multiple exposures to be made, which enables optimization of signal to noise. 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. Chemiluminescent detection for western blotting is popular because it offers several advantages over other detection methods. 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. 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. Chemiluminescent Western blots are popular assays for assessing protein expression. 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. 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 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). ![]()
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