Thaw the ladder either at room temperature or at 37-40☌ for a few minutes to dissolve precipitated solids. The quality of the BLUeye Prestained Protein Ladder is tested on a lot-to-lot basis to ensure consistent product quality.īLUeye Prestained Protein Ladder Protocolġ. High-affinity binding of StrepTactin, a genetically modified form of streptavidin, gives convenient and simultaneous detection of both proteins and standards on western blots using either colorimetric or chemiluminescence methods. Sizing of proteins on SDS-PAGE and Western blots.Īpproximately 0.1~0.4 mg/ml of each protein in the 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 I used your Thermo Scientific Unstained Protein MW Marker for a western transfer and got very poor transfer onto the membrane. ![]() The protein standard is supplied in a ready-to-use format for direct loading onto gels no need to heat, reduce, or add sample buffer prior to use. The ladder is supplied in gel loading buffer and is ready to use.īroad range: 10-245 kDa (Tris-glycine-SDS running buffer) Add freshly prepared DTT solution to a final concentration of 50 mM. For detection of any western blot, it is desirable to use prestained molecular weight markers (also called protein ladders) that are transferred to the membrane along with the protein sample. SeeBlue Plus2 Pre-Stained Standard contains 10 proteins (4250 kDa): 8 blue-dyed and 2 with contrasting colors, for easy and quick evaluation of electrophoresis and western transfer efficiency. The BLUeye Prestained Protein Ladder is designed for monitoring protein separation during, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size 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. ![]() My experimental protein size is 124kDA and I choose Beta Actin as a loading control which is small. The BLUeye Prestained Protein Ladder is a three-color protein standard with 12 prestained proteins covering a wide range molecular weights from 10 to 245 kilodalton (kDa). I am a beginner of western blot and from some days I have been facing a lot of problems.
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