10 μl per well for large gels, 5 μl per well for blots.5 μl per well for mini-gels, 2.5 μl per well for blots.Load the following volumes of the ladder on SDS-PAGE gel:
Mix thoroughly to ensure the solution is homogeneous.ģ. Thaw the ladder either at room temperature or at 37-40☌ for a few minutes to dissolve precipitated solids. Do not boil.Ģ. The quality of the BLUelf Prestained Protein Ladder is tested on a lot-to-lot basis to ensure consistent product quality.īLUelf Prestained Protein Ladder Protocolġ. Sizing of proteins on SDS-PAGE and Western blots.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 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.Easy to identify: includes the ~25, ~75 kDa reference bands coupled with a green and a red dye.Ready-to-use: supplied in a loading buffer for direct loading on gels.Broad range: 3.5-245 kDa (Tris-glycine-SDS running buffer).Do not heats, dilute, and add reducing agent before loading. The ladder is supplied in gel loading buffer and is ready to use. The BLUelf Prestained Protein Ladder is designed for monitoring protein separation during SDS-PAGE, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of p roteins. 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 BLUelf Prestained Protein Ladder is a three-color protein standard with 13 prestained proteins covering a wide range molecular weights from 3.5 to 245 kilodalton (kDa).